VOLUME 9 • MARCH 1989 • NUMBER 3
`
`Published monthly by the
`American Society for Microbiology
`
`9
`
`(3)
`
`875-1380
`
`(1989)
`
`ISSN : 0270-7306
`
`BIOEPIS EX. 1021
`Page 1
`
`

`

`MOLECULAR AND CELLULAR BIOLOGY
`
`VOLUME 9 • MARCH 1989 • NUMBER 3
`
`Aaron J. Shatkin, Editor in Chief(1990)
`.,Center for Advanced
`Biotechnology and Medicine
`Piscataway, N.J.
`Tony Hunter, Editor (1993)
`The Salk Institute
`San Diego, Calif.
`David J. L. Luck, Editor (1992)
`Rockefeller University
`New York, N.Y.
`Steven L. McKnight, Editor (1992)
`Carnegie Institution of Washington
`Baltimore, Md.
`
`Randy W. Schekman, Editor
`(1992)
`University of California
`Berkeley
`Louis Siminovitch, Editor (1990)
`Mount Sinai Hospital
`Toronto, Canada
`Joan A. Steitz, Editor (1990)
`Yale University
`New Haven, Conn.
`
`Shirley M. Tilghman, Editor (1993)
`Princeton University
`Princeton, N.J.
`Robert Tjian, Editor (1991)
`University of California
`Berkeley
`Alan M. Weiner, Editor (1993)
`Yale University
`New Haven, Conn.
`Owen N. Witte, Editor (1993)
`Molecular Biology Institute
`University of California
`Los Angeles
`
`Frederick W. Alt (1990)
`Susan Berget (1990)
`Arnold J. Berk (1991)
`Alan Bernstein (1990)
`Barbara K. Birshtein (1990)
`J. Michael Bishop (1990)
`Elizabeth H. Blackburn
`(1991)
`Jef D. Boeke (1991)
`Michael R. Botchan (1990)
`David Botstein (1990)
`Bruce P. Brandhorst (1990)
`Marjorie C. Brandriss
`(1991)
`Kathryn Calame (1991)
`Mario R. Capecchi (1990)
`John A. Carbon (1990)
`Marian Carlson (1989)
`C. Thomas Caskey (1990)
`Lawrence A. Chasin (1991)
`Nam-Hai Chua (1991)
`Don W. Cleveland (1990)
`Nicholas Cowan (1990)
`Elizabeth A. Craig (1991)
`Thomas Curran (1991)
`Mark Davis (1990)
`Gideon Dreyfuss (1990)
`
`EDITORIAL BOARD
`Mary P. Edmonds (1990)
`Marilyn Kozak (1991)
`Robert N. Eisenman (1991)
`Monty Krieger (1989)
`Sarah C. R. Elgin (1991)
`Elizabeth Lacy (1990)
`Scott Emr (1991)
`Alan Lambowitz (1990)
`Walton L. Fangman (1991)
`Michael Lenardo (1991)
`Thomas D. Fox (1990)
`Arthur D. Levinson (1990)
`Mary-Jane Gething (1990)
`Susan Lindquist (1990)
`Steve Goff (1989)
`Stuart M. Linn (1989)
`Jack F. Greenblatt (1991)
`Douglas Lowy (1990)
`Leonard P. Guarente (1991)
`Paul T. Magee (1991)
`Christine Guthrie (1989)
`James Manley (1989)
`James E. Haber (1990)
`Kunihiro Matsumoto (1991)
`Hidesaburo Hanafusa (1989)
`Janet E. Mertz (1990)
`Bernardo Nadai-Ginard (1990)
`Leland D. HartweU (1990)
`Nathaniel Heintz (1991)
`Paul Neiman (1989)
`Ari Helenius (1990)
`Joseph R. Nevins (1990)
`Steven Henikoff (1991)
`Carol Newlon (1991)
`Ira Herskowitz (1990)
`Mary Ann Osley (1990)
`James B. Hicks (1989)
`Haney L. Ozer (1991)
`Alan Hinnebusch (1991)
`Mary Lou Pardue (1991)
`Michael J. HoUand (1990)
`David Patterson (1991)
`Greg HoUis (1990)
`Gianni Piperno (1991)
`Anita K. Hopper (1991)
`John R. Pringle (1991)
`Peter M. Howley (1991)
`Steven I. Reed (1991)
`Mark Johnston (1991)
`Jean-Paul Revel (1991)
`Daniel Klessig (1989)
`Jasper D. Rine (1991)
`Barbara Knowles (1989)
`Naomi E. Rosenberg (1991)
`
`Rodney Rothstein (1990)
`Norman P. Salzman (1991)
`Paul Schedl (1990)
`Milton J. Schlesinger (1989)
`Matthew P. Scott (1989)
`Charles J. Sherr (1991)
`Arthur Skoultchi (1991)
`Barbara SoOner-Webb (1989)
`Frank Solomon (1991)
`Michael B. Sporn (1991)
`Allan C. Spradling (1991)
`Karen Sprague (1989)
`Pamela Stanley (1991)
`Nat Sternberg (1989)
`Bruce Stillman (1991)
`Kevin Struhl (1989)
`Kenneth D. Stuart (1991)
`Bill Sugden (1991)
`Lawrence H. Thompson (1991)
`Geoffrey Wahl (1989)
`Peter Walter (1990)
`Jouathan R. Warner (1990)
`Reed B. Wickner (1991)
`Lewis T. Williams (1991)
`Fred Winston (1991)
`Elton T. Young (1990)
`Michael Young (1991)
`
`Helen R. Whiteley, Chairman, Publications Board
`Linda M. Illig, Managing Editor, Journals
`
`Kirk Jensen, Director of Publications
`Linda M. DUg, Production Editor
`
`Molecular and Cellular Biology (ISSN 0270-7306), a publication of the American Society for Microbiology, 1913 I St., N. W.,
`Washington, DC 20006-5107, is devoted to the advancement and dissemination of fundamental knowledge concerning the
`molecular biology of eucaryotic cells, of both microbial and higher organisms. Instructions to authors are published in the
`January issue each year; reprints are available from the editors and the Publications Department. The journal is published
`monthly, one volume per year. The nonmember subscription price is $340 per year; single copies are $30. The member
`subscription price is $43 (foreign, $71 [air drop shipping]) per year; single copies are $8. Correspondence relating to
`subscriptions, reprints, defective copies, availability of back issues, lost or late proofs, disposition of submitted manuscripts,
`and general editorial matters should be directed to the ASM Publications Department, 1913 I St., N.W., Washington, DC
`20006-5107 (area 202 833-9680).
`Claims for missing issues from residents of the United States, Canada, and Mexico must be submitted within 3 months after
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`POSTMASTER: Send address changes to Molecular and Cellular Biology, ASM, 1913 I St., N. W., Washington, DC 20006-
`5107.
`Made in the United States of America. Printed on acid-free paper.
`Copyright© 1989, American Society for Microbiology.
`All Rights Reserved.
`The code at the top of the first page of an article in this journal indicates the copyright owner's consent that copies of the article
`may be made for personal use or for personal use of specific clients. This consent is given on the condition, however, that the
`copier pay the stated per-copy fee through the Copyright Clearance Center Inc., 21 Congress St., Salem, MA 01970, for
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`works, or for resale.
`
`B*: flim:.HH~J<,J< l: :t.lll!f#.'..: < l'ltt,
`
`BIOEPIS EX. 1021
`Page 2
`
`

`

`Agabian, Nina, 1212
`Alitalo, Kari, 1255
`Arceci, Robert J., 1346
`
`Barringer, Kevin J., 1316
`Bedard, Pierre-Andre, 1371
`Bedwell, David M., 1014
`Beeman, Karen, 1155
`Bernstein, Mitchell, 1191
`Bertrand, Helmut, 1362
`Birshtein, Barbara K., 1324
`Bishop, J. Michael, 1148
`Bissinger, Peter H., 1309
`Bloom, Kerry, 1368
`Bos, Timothy J., 1255
`Brownlee, Clare, 1060
`Brugge, Joan S., 1109
`Burke, Dan, 1049
`
`Calos, Michele P., 1026
`Carattoli, Alessandra, 1271
`Carle, GeorgeS., 983
`Chae, Chi-Born, 1005
`Chalkley, Roger, 1289
`Challoner, Peter B., 902
`Chedid, Marcia, 959
`Chin, William W., 1128
`Clayton, Christine E., 1332
`Clayton, David A., 1200
`Conscience, Jean-Franc;ois,
`1183
`Cregg, James M., 1316
`Croop, James M., 1346
`Culotta, Valeria Cizewski,
`1376
`
`Darling, Douglas S., 1128
`Davies, Monique V., 946,
`1233
`DeMarco, Michael, 1109
`Deuring, Renate, 875
`Deutsch, Walter A., 965
`Devault, Alain, 1346
`Diamantis, Ioannis D., 1183
`Didion, Thomas, 988
`Dihanich, Melitta, 1100
`Dorner, Andrew J., 1233
`Dubin, Robert A., 1083
`
`Ehrlich, Kenneth C., 1351
`Ehrlich, Melanie, 1351
`Elliott, David J., 1069
`Emr, Scott D., 1014
`Erikson, R. L., 1371
`
`Felsenfeld, Gary, 893
`Fendly, Brian M., 1165
`Fisch, Tobe M., 1327
`Fluck, Michele M., 995
`Foreman, Pamela K., 1137
`Fox, Larry G., 1109
`Frankel, Wayne, 1284
`Fu, Ying-Hui, 1120
`Fuller, Margaret T., 875
`
`Author Index
`
`Garcia-Blanco, Mariano A.,
`1060
`Gasdaska, Pam, 1049
`Ghazal, Peter, 1342
`Granner, Daryl K., 1289
`Groen, N. A., 1277
`Gras, Philippe, 1346
`Groudine, Mark, 902
`
`Haase, Steven B., 1026
`Haber, Daniel, 1346
`Hacker, David, 995
`Hamer, Dean H., 1376
`Hamilton, Barbara, 1309
`Hamlin, Joyce L., 1137
`Hanahan, Douglas, 925
`Hannigan, Gregory, 1060
`Harless, Julie, %5
`Hartwell, Leland, 1049
`Hays, Thomas S., 875
`Hennighausen, Lothar, 1342
`Hershey, John W. B., 946
`Hill, Alison, 1368
`Hodin, Richard A., 1128
`Hofer, Paul, 1183
`Holland, Janice P., 1243
`Holland, Michael J., 1243
`Housman, David E., 1346
`Howley, Peter M., 925
`Hudziak, Robert M., 1165
`Hwang, Inhwan, 1005
`
`lp, Y. Tony, 1289
`Irving, Steven G., 1034, 1041
`Israel, David 1., 1233
`
`Jacobs, Howard T., 1069
`Jerry, D. Joseph, 935
`Johnston, Leslie A., 935
`Jones, Peter A., 885
`Jongeward, Gregg D., 1014
`June, Carl H., 1034
`
`Kang, John, 1243
`Katzen berg, Daniel R., 1324
`Kaufman, Randal J., 946,
`1233,
`Kelley, Mark R., %5
`Kelly, Kathleen, 1034, 1041
`Kepes, Franc;ois, 1191
`Keski-Oja, Jorma, 1255
`Kim, Ki-Han, 974
`Kindler, Vincent, 1183
`Knezetic, Joseph A., 893
`Kobayashi, George S., 983
`Kotarski, Michael A., 935
`Kowallik, Klaus V., 988
`Kozak, Leslie P., 935
`Krag, Sharon S., 914
`Krysan, Patrick J., 1026
`
`Lambowitz, Alan M., 1362
`Lambris, John D., 1100
`Lazar, Mitchell A., 1128
`
`Lengyel, Peter, 1060
`Lewis, Gail D., 1165
`Ling, Victor, 1224
`Lizardi, Paul M., 1212
`Lockshin, Curtis, 1342
`Lohman, P. H. M., 1277
`Lowenhaupt, Ky, 1173
`Lubon, Henryk, 1342
`Lugo, Tracy Gross, 1263
`Liihrmann, Reinhard, 1212
`
`Macino, Giuseppe, 1271
`Madden, Knut R., 1316
`Mannix, Daniel G., 1120
`Marshallsay, Brigitte, 1100
`Marzluf, George A., 1120
`Mcintosh, Lee, 1362
`Medoff, Gerald, 983
`Mestel, Rosie, 1243
`Michalowsky, Lesley A., 885
`Mizel, Steven B., 959
`Morelli, Giorgio, 1271
`Moroni, Christoph, 1183
`Morrison, Elizabeth, 1060
`Moss, Stuart B., 902
`Mottram, Jeremy, 1212
`Mowatt, Michael R., 1332
`
`Nair, Asha P. K., 1183
`Nelson, F. Kenneth, 1284
`Nelson, Jay, 1342
`Nelson, Mary Anne, 1271
`Nelson, Richard G., 1212
`Nemeth, Susan P., 1109
`Neumann, Dieter, 1298
`Ng, William F., 1224
`Nickels, Roxy, 1362
`Nover, Lutz, 1298
`Nusse, Roe!, 1357
`
`Paietta, John V., 1120
`Pape, Michael E., 974
`Pardue, Mary Lou, 1173
`Pathak, Vinay K .. 946
`Perry, Karen L., 1212
`Pertovaara, Liisa, 1255
`Piatigorsky, Joram, 1083
`Pollok, Brian A., 959
`Prout, Mary, 875
`Prywes, Ron, 1327
`
`Rajan, T.V .. 1284
`Ray, DanS., 1365
`Raymond, Martine, 1346
`Reynolds-Kohler, Catherine,
`1342
`Rich, Alexander, 1173
`Robertson, Barbara, 875
`Roeder, Robert G., 1327
`Roelink, Henk, 1357
`Roggenkamp, Rainer, 988
`Romano, Nicoletta, 1271
`Rosenwald, Anne G., 914
`Ruis, Helmut, 1309
`
`Rutherford, Michael, 1060
`Ryden, Thomas A., 1155
`
`Sabourin, Josanne R., 1362
`Sarangi, Farida, 1224
`Scharf, Klaus-Dieter, 1298
`Schekman, Randy, 1191
`Schuuring, Ed, 1357
`Shepard, H. Michael, 1165
`Shirakawa, Fumihiko, 959
`Shtivelman, E., 1148
`Siebenlist, Ulrich, 1034, 1041
`Simmons, Daniel L., 1371
`Simons, J. W. I. M., 1277
`Sippola-Thiele, Maria, 925
`Sistonen, Lea, 1255
`Stanley, Pamela, 914
`Steele, Paul E., 983
`Stiles, Charles D., 1060
`Stillman, Cathy A., 1316
`Strobel, Scott A., 1014
`Supakar, Prakash C., 1351
`Suttles, Jill, 959
`
`Thill, Gregory P., 1316
`Tilley, Shermaine A., 1324
`Topper, James N., 1200
`
`Ullrich, Axel, 1165
`
`van Deemter, Liesbeth, 1357
`van Rooijen, M. L., 1277
`Van Tuinen, Evert, 1100
`van Zeeland, A. A., 1277
`Veinot-Drebot, Lela, 1224
`Venugopal, Sheela, 965
`Verma, Ioder M., 1336
`Visvader, Jane, 1336
`Vogt, Peter K., 1255
`Vrieling, H., 1277
`
`Wang, Enduo, 1243
`Wasley, Louise C., 1233
`Wawrousek, Eric F., 1083
`Wieser, Rotraud, 1309
`Williams, Bryan R. G., 1060
`Winget, Marcy, 1165
`Wisdom, Gregory S., 1332
`Wise, Robert J., 1233
`Witte, Owen N., 1263
`
`Yannoni, Yvonne, 1371
`Yao, Ching-Ho, 1092
`Yao, Meng-Chao, 1092
`Yip, Michele, 1243
`Yun, Kyuson, 1014
`
`Zastawny, Roman L., 1224
`Zdzienicka, M. Z., 1277
`Zhang, Daoling, 1351
`Zipfel, Peter F., 1034, 1041
`
`BIOEPIS EX. 1021
`Page 3
`
`

`

`MOLECULAR AND CELLULAR BIOLOGY
`
`Volume 9
`
`March 1989
`
`No.3
`
`GENE EXPRESSION
`
`Gene Structure and Transcription in Mouse Cells with Extensively Demethylated
`DNA. Lesley A. Michalowsky and Peter A. Jones......................
`Identification and Characterization of a Chicken a-Globin Enhancer. Joseph A.
`Knezetic and Gary Felsenfeld . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`Control of Carbohydrate Processing: Increased (.l-1,6 Branching inN-Linked
`Carbohydrates of Lec9 CHO Mutants Appears To Arise from a Defect in
`Oligosaccharide-Dolichol Biosynthesis. Anne G. Rosenwald, Pamela
`Stanley, and Sharon S. Krag . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`An Ubiquitously Expressed Gene 3.5 Kilobases Upstream of the Glycerol-3-
`Phosphate Dehydrogenase Gene in Mice. Leslie A. Johnston, Michael A.
`Kotarski, D. Joseph Jerry, and Leslie P. Kozak.........................
`The Phosphorylation State of Eucaryotic Initiation Factor 2 Alters Translational
`Efficiency of Specific mRNAs. Randal J. Kaufman, Monique V. Davies,
`Vinay K. Pathak, and John W. B. Hershey.............................
`Interleukin 1 and Cyclic AMP Induce K Immunoglobulin Light-Chain Expression
`via Activation of an NF-KB-Like DNA-Binding Protein. Fumihiko
`Shirakawa, Marcio Chedid, Jill Suttles, Brian A. Pollok, and Steven B.
`Mizel . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`Transcriptional Regulation of Acetyl Coenzyme A Carboxylase Gene Expression
`by Tumor Necrosis Factor in 30A-5 Preadipocytes. Michael E. Pape and
`Ki-Han Kim.........................................................
`S-Phase-Specific Transcription Regulatory Elements Are Present in a Replication(cid:173)
`Inhwan Hwang and
`Independent Testis-Specific H2B Histone Gene.
`Chi-Born Chae.......................................................
`Mitogen-Induced Genes Are Subject to Multiple Pathways of Regulation in the
`Initial Stages ofT-Cell Activation. Steven G. Irving, Carl H. June, Peter
`F. Zipfel, Ulrich Siebenlist, and Kathleen Kelly.........................
`Complexity of the Primary Genetic Response to Mitogenic Activation of Human T
`Cells. Peter F. Zipfel, Steven G. Irving, Kathleen Kelly, and Ulrich
`Siebenlist . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`Regulation of 2' ,5' -Oligoadenylate Synthetase Gene Expression by Interferons and
`Platelet-Derived Growth Factor. Mariano A. Garcia-Blanco, Peter
`Lengyel, Elizabeth Morrison, Clare Brownlee, Charles D. Stiles, Michael
`Rutherford, Gregory Hannigan, and Bryan R. G. Williams . . . . . . . . . . . . . . .
`Mutually Exclusive Synthetic Pathways for Sea Urchin Mitochondrial rRNA and
`mRNA. David J. Elliott and Howard T. Jacobs . . . . . . . . . . . . . . . . . . . . . . . .
`Expression of the Murine aD-Crystallin Gene Is Not Restricted to the Lens.
`Robert A. Dubin, Eric F. Wawrousek, and Joram Piatigorsky . . . . . . . . . . . .
`cys-3, the Positive-Acting Sulfur Regulatory Gene of Neurospora crassa, Encodes
`a Protein with a Putative Leucine Zipper DNA-Binding Element. Ying-Hui
`Fu, John V. Paietta, Daniel G. Mannix, and George A. Marzluf...........
`A Novel Member of the Thyroid/Steroid Hormone Receptor Family Is Encoded
`by the Opposite Strand of the Rat c-erbAa Transcriptional Unit. Mitchell
`A. Lazar, Richard A. Hodin, DouglasS. Darling, and William W. Chin....
`
`885-892
`
`893-901
`
`914-924
`
`935-945
`
`946--958
`
`959-964
`
`974-982
`
`1005-1013
`
`1034-1040
`
`1041-1048
`
`1060-1068
`
`1069-1082
`
`1083-1091
`
`1120-1127
`
`1128-1136
`
`Continued on following page
`
`BIOEPIS EX. 1021
`Page 4
`
`

`

`Continued from preceding puge
`
`Avian Retroviral Long Terminal Repeats Bind CCAAT/Enhancer-Binding
`Protein. Thomas A. Ryden and Karen Beemon . . . . . . . . . . . . . . . . . . . . . . . .
`Identification of Transcriptional Regulatory Elements in Human Mitochondrial
`DNA by Linker Substitution Analysis. James N. Topper and David A.
`Clayton. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`Isolation and Sequence of Four Small Nuclear U RNA Genes of Trypanosoma
`brucei subsp. brucei: Identification of the U2, U4, and U6 RNA Analogs.
`Jeremy Mottram, Karen L. Perry, Paul M. Lizardi, Reinhard LOhrmann,
`Nina Agabian, and Richard G. Nelson . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`Effect of von Willebrand Factor Coexpression on the Synthesis and Secretion of
`Factor VIII in Chinese Hamster Ovary Cells. Randal J. Kaufman, Louise
`C. Wasley, Monique V. Davies, Robert J. Wise, David I. Israel, and
`Andrew J. Domer....................................................
`Sequences within the Spacer Region of Yeast rRNA Cistrons That Stimulate 35S
`rRNA Synthesis In Vivo Mediate RNA Polymerase 1-Dependent Promoter
`and Terminator Activities. Rosie Mestel, Michele Yip, Janice P. Holland,
`Enduo Wang, John Kang, and Michael J. Holland. . . . . . . . . . . . . . . . . . . . . . .
`Enhanced jun Gene Expression Is an Early Genomic Response to Transforming
`Growth Factor 13 Stimulation. Liisa Pertovaara, Lea Sistonen, Timothy J.
`Bos, Peter K. Vogt, Jorma Keski-Oja, and Kari Alitalo..................
`The BCR-ABL Oncogene Transforms Rat-1 Cells and Cooperates with v-myc.
`Tracy Gross Lugo and Owen N. Witte.................................
`Molecular Cloning of a Neurospora crassa Carotenoid Biosynthetic Gene
`(Aibino-3) Regulated by Blue Light and the Products of the White Collar
`Genes. Mary Anne Nelson, Giorgio Morelli, Alessandra Carattoli,
`Nicoletta Romano and Giuseppe Macino . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`Hormonal Regulation of Phosphoenolpyruvate Carboxykinase Gene Expression Is
`Mediated through Modulation of an Already Disrupted Chromatin
`Structure. Y. Tony lp, Daryl K. Granner, and Roger Chalkley . . . . . . . . . .
`Cytoplasmic Heat Shock Granules Are Formed from Precursor Particles and Are
`Associated with a Specific Set of mRNAs. Lutz Nover, Klaus-Dieter
`Scharf, and Dieter Neumann..........................................
`Functional Characterization of the Two Alcohol Oxidase Genes from the Yeast
`James M. Cregg, Knut R. Madden, Kevin J. Barringer,
`Pichia pastoris.
`Gregory P. Thill, and Cathy A. Stillman................................
`Nucleotide Sequence of an Unequal Sister Chromatid Exchange Site in a Mouse
`Myeloma Cell Line. Daniel R. Katzen berg, Shermaine A. Tilley, and
`Barbara K. Birshtein.................................................
`An APt-Binding Site in the c-fos Gene Can Mediate Induction by Epidermal
`Growth Factor and 12-0-Tetradecanoyl Phorbol-13-Acetate. Tobe M.
`Fisch, Ron Prywes, and Robert G. Roeder . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`Differential Transcription of Exon 1 of the Human c-fms Gene in Placental
`Trophoblasts and Monocytes. Jane Visvader and Inder M. Verma........
`Cell-Specific Activity of the Modulator Region in the Human Cytomegalovirus
`Major Immediate-Early Gene. Henryk Lubon, Peter Ghazal, Lothar
`Henriighausen, Catherine Reynolds-Kohler, Curtis Lockshin, and Jay
`Nelson. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`The Three Mouse Multidrug Resistance (mdr) Genes Are Expressed in a Tissue(cid:173)
`Specific Manner in Normal Mouse Tissues. James M. Croop, Martine
`Raymond, Daniel Haber, Alain Devault, Robert J. Arceci, Philippe Gros,
`and David E. Housman.. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`
`1155-1164
`
`1200-1211
`
`1212-1223
`
`1233-1242
`
`1243-1254
`
`1255-1262
`
`1263-1270
`
`1271-1276
`
`1289-1297
`
`1298-1308
`
`1316--1323
`
`1324-1326
`
`1327-1331
`
`1336--1341
`
`1342-1345
`
`1346--1350
`
`Continued on following page
`
`BIOEPIS EX. 1021
`Page 5
`
`

`

`Continued from preceding page
`
`Rapid Repression of Quiescence-Specific Gene Expression by Epidermal Growth
`Factor, Insulin, and pp6ov-src. Pierre-Andre Bedard, Yvonne Yannoni,
`Daniel L. Simmons, and R. L. Erikson. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`Fine Mapping of a Mouse Metallothionein Gene Metal Response Element.
`Valeria Cizewski Culotta and Dean H. Hamer . . . . . . . . . . . . . . . . . . . . . . . . . .
`
`1371-1375
`
`1376-1380
`
`CELL GROWTH AND DEVELOPMENT
`Expression of Replication-Dependent Histone Genes in Avian Spermatids Involves
`an Alternate Pathway of mRNA 3'-End Formation. Peter B. Challoner,
`Stuart B. Moss, and Mark Groudine...................................
`Cell-Heritable Stages of Tumor Progression in Transgenic Mice Harboring the
`Bovine Papillomavirus Type 1 Genome. Maria Sippola-Thiele, Douglas
`Hanahan, and Peter M. Howley.......................................
`Dominant Effects of Tubulin Overexpression in Saccharomyces cerevisiae. Dan
`Burke, Pam Gasdaska, and Leland Hartwell............................
`Deletions within the Amino-Terminal Half of the c-src Gene Product That Alter
`the Functional Activity of the Protein. Susan P. Nemeth, Larry G. Fox,
`Michael DeMarco, and Joan S. Brugge. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`The PVT Gene Frequently Amplifies with MYC in Tumor Cells. E. Shtivelman
`and J. Michael Bishop................................................
`pl85HER2 Monoclonal Antibody Has Antiproliferative Effects In Vitro and
`Sensitizes Human Breast Tumor Cells to Tumor Necrosis Factor. Robert
`M. Hudziak, Gail D. Lewis, Marcy Winget, Brian M. Fendly, H. Michael
`Shepard, and Axel Ullrich . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`A v-H-ras-Dependent Hemopoietic Tumor Model Involving Progression from a
`Clonal Stage of Transformation Competence to Autocrine Interleukin 3
`Production. Asha P. K. Nair, Ioannis D. Diamantis, Jean-Fran<;ois
`Conscience, Vincent Kindler, Paul Hofer, and Christoph Moroni..........
`Control of Saccharomyces cerevisiae Catalase T Gene (CTTJ) Expression by
`Nutrient Supply via the RAS-Cyclic AMP Pathway. Peter H. Bissinger,
`Rotraud Wieser, Barbara Hamilton, and Helmut Ruis....................
`Variation of Tandem Repeats in the Developmentally Regulated Procyclic Acidic
`Repetitive Proteins of Trypanosoma brucei. Michael R. Mowatt, Gregory S.
`Wisdom, and Christine E. Clayton. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`Transient Expression of the Proto-Oncogene int-1 during Differentiation of P19
`Embryonal Carcinoma Cells. Ed Schuuring, Liesbeth van Deemter, Henk
`Roelink, and Roel Nusse . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`
`CELL AND ORGANELLE STRUCTURE AND ASSEMBLY
`Interacting Proteins Identified by Genetic Interactions: a Missense Mutation in
`a-Tubulin Fails To Complement Alleles of the Testis-Specific P· Tubulin Gene
`of Drosophila melanogaster. Thomas S. Hays, Renate Deuring, Barbara
`Robertson, Mary Prout, and Margaret T. Fuller ........................ .
`Formation of Irregular Giant Peroxisomes by Overproduction of the Crystalloid
`Core Protein Methanol Oxidase in the Methylotrophic Yeast Hansenula
`polymorpha. Rainer Roggenkamp, Thomas Didion, and Klaus V.
`Kowallik ........................................................... .
`
`902-913
`
`925-934
`
`1049-1059
`
`1109-1119
`
`1148-1154
`
`1165-1172
`
`1183-1190
`
`1309-1315
`
`1332-1335
`
`1357-1361
`
`875-884
`
`988-994
`
`Continued on following page
`
`BIOEPIS EX. 1021
`Page 6
`
`

`

`Continued from preceding page
`
`Sequence and Structural Requirements of a Mitochondrial Protein Import Signal
`Defined by Saturation Cassette Mutagenesis. David M. Bedwell, Scott A.
`Strobel, Kyuson Yon, Gregg D. Jongeward, and Scott D. Emr. . . . . . . . . . . .
`Accumulation of Viruslike Particles in a Yeast Mutant Lacking a Mitochondrial
`Pore Protein. Melitta Dihanich, Evert Van Tuinen, John D. Lambris, and
`Brigitte Marshallsay..................................................
`SEC59 Encodes a Membrane Protein Required for Core Glycosylation in
`Saccharomyces cerevisiae. Mitchell Bernstein, Fran~ois Kepes, and Randy
`Schekman. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`Immunological Identification of the Alternative Oxidase of Neurospora crassa
`Mitochondria. Alan M. Lambowitz, Josanne R. Sabourin, Helmut
`Bertrand, Roxy Nickels, and Lee Mcintosh.............................
`Conserved Sequence Blocks in Kinetoplast Minicircles from Diverse Species of
`Trypanosomes. DanS. Ray..........................................
`
`1014-1025
`
`1100-1108
`
`1191-1199
`
`1362-1364
`
`1365-1367
`
`CHROMOSOME STRUCTURE AND DYNAMICS
`Antibody to a Human DNA Repair Protein Allows for Cloning of a Drosophila
`eDNA That Encodes an Apurinic Endonuclease. Mark R. Kelley, Sheela
`Venugopal, Julie Harless, and Walter A. Deutsch . . . . . . . . . . . . . . . . . . . . . . .
`Electrophoretic Analysis of Histoplasma capsulatum Chromosomal DNA. Paul E.
`Steele, Georges F. Carle, GeorgeS. Kobayashi, and Gerald Medoff.......
`High-Level Recombination Specific to Polyomavirus Genomes Targeted to the
`Integration-Transformation Pathway. David Hacker and Michele M. Fluck
`Isolation of Human Sequences That Replicate Autonomously in Human Cells.
`Patrick J. Krysan, Steven B. Haase, and Michele P. Calos...............
`Accurate Processing and Amplification of Cloned Germ Line Copies of Ribosomal
`DNA Injected into Developing Nuclei of Tetrahymena thermophila. Meng-
`Chao Yao and Ching-Ho Yao . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`Identification and Characterization of a Gene That Is Coamplified with
`Dihydrofolate Reductase in a Methotrexate-Resistant CHO Cell Line.
`Pamela K. Foreman and Joyce L. Hamlin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`Nonrandom Distribution of Long Mono- and Dinucleotide Repeats in Drosophila
`Chromosomes: Correlations with Dosage Compensation, Heterochromatin,
`and Recombination. Ky Lowenhaupt, Alexander Rich, and Mary Lou
`Pardue..............................................................
`Identification of Members of the P-Giycoprotein Multigene Family. William F.
`Ng, Farida Sarangi, Roman L. Zastawny, Lela Veinot-Drebot, and Victor
`Ling................................................................
`DNA Strand Specificity for UV-Induced Mutations in Mammalian Cells. H.
`Vrieling, M. L. van Rooijen, N. A. Groen, M. Z. Zdzienicka, J. W. I. M.
`Simons, P. H. M. Lohman, and A. A. van Zeeland......................
`Mitotic Recombination Is Responsible for the Loss of Heterozygosity in Cultured
`Murine Cell Lines. F. Kenneth Nelson, Wayne Frankel, and T.V. Rajan
`A Plant DNA-Binding Protein That Recognizes 5-Methylcytosine Residues.
`Daoling Zhang, Kenneth C. Ehrlich, Prakash C. Supakar, and Melanie
`Ehrlich . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`Acquisition and Processing of a Conditional Dicentric Chromosome in
`Saccharomyces cerevisiae. Alison Hill and Kerry Bloom . . . . . . . . . . . . . . . . .
`
`%5-973
`
`983-987
`
`995-1004
`
`1026-1033
`
`1092-1099
`
`1137-1147
`
`1173-1182
`
`1224-1232
`
`1277-1283
`
`1284-1288
`
`1351-1356
`
`1368-1370
`
`Date of Issue: 24 February 1989
`
`BIOEPIS EX. 1021
`Page 7
`
`

`

`MOLECULAR AND CELLULAR BIOLOGY, Mar. 1989, p. 1165-1172
`0270-7306/89/031165-08$02.00/0
`Copyright © 1989, American Society for Microbiology
`
`Vol. 9, No.3
`
`p185HER2 Monoclonal Antibody Has Antiproliferative Effects In
`Vitro and Sensitizes Human Breast Tumor Cells to
`Tumor Necrosis Factor
`ROBERT M. HUDZIAK,1 GAIL D. LEWIS,2 MARCY WINGET,3 BRIAN M. FENDLY,3 H. MICHAEL SHEPARD,2
`AND AXEL ULLRICH 1t*
`Departments of Developmental Biology, 1 Pharmacological Sciences, 2 and Medicinal and Analytical Chemistry, 3
`Genentech, Inc., 460 Point San Bruno Boulevard, South San Francisco, California 94080
`
`Received 3 October 1988/Accepted 8 December 1988
`
`The HERl/c-erbB-2 gene encodes the epidermal growth factor receptorlike human homolog of the rat neu
`oncogene. Amplification of this gene in primary breast carcinomas has been shown to correlate with poor
`clinical prognosis for certain ~r patients. We show here that a monoclonal antibody directed against the
`extraceUular domain of p185HER2 specifically inhibits the growth of breast tumor-derived ceO lines overex(cid:173)
`pressing the HERl/c-erbB-2 gene product and prevents HERl/c-erbB-2-traQsformed NIH 3T3 cells from
`forming colonies in soft agar. Furthermore, resistance to the cytotoxic effect of tumor necrosis factor alpha,
`which has been shown to be a consequence of HERl/c-erbB-2 overexpression, is significantly reduced in the
`presence of this antibody.
`
`HER2/c-erbB-2, the human homolog of the rat proto(cid:173)
`oncogene neu (4, 34), encodes a 1,255-amino-acid glycopro(cid:173)
`tein with extensive homology to the human epidermal
`growth factor (EGF) receptor (4, 21, 33, 34, 42). The
`HER2/c-erbB-2 gene product, p185HER2 , has all of the struc(cid:173)
`tural features and many of the functional properties of
`subclass I growth factor receptors (reviewed in references 43
`and 44), including cell surface location and an intrinsic
`tyrosine kinase activity. However, the ligand for this puta(cid:173)
`tive growth factor receptor has not yet been identified.
`Amplification of the HER2/c-erbB-2 gene has been found
`in human salivary gland and gastric tumor-derived cell lines
`(13, 34), as well as in mammary gland carcinomas (21, 22, 40,
`42). Slamon et al. (35) surveyed 189 primary breast adeno(cid:173)
`carcinomas and determined that the HER2/c-erbB-2 gene
`was amplified in about 30% of the cases. Most importantly,
`HER2/c-erbB-2 amplification was correlated with a negative
`prognosis and high probability of relapse. Similar although
`less frequent amplification of the HER21c-erbB-2 gene has
`been reported for gastric and colon adenocarcinomas (45,
`46). Experiments with NIH 3T3 cells also suggest a direct
`role for the overexpressed, structurally unaltered HER21
`c-erbB-2 gene product p185HER2 in neoplastic transforma(cid:173)
`tion. High levels of HER2/c-erbB-2 gene expression attained
`by coamplification of the introduced gene with dihydrofolate
`reductase by methotrexate selection (18) or by using a strong
`promoter (6) was shown to transform NIH 3T3 fibroblasts.
`Only cells with high levels ofp185HER2 are transformed, i.e.,
`have an altered morphology, are anchorage independent,
`and will form tumors in athymic mice.
`Overexpression ofp185HER2 may, furthermore, contribute
`to malignant tumor development by allowing tumor cells to
`evade one component of the antitumor defenses of the body,
`the activated macrophage (17). Macrophages play an impor(cid:173)
`tant role in immune surveillance against neoplastic growth in
`vivo (1, 2, 38), and Urban et al. (39) have shown that tumor
`
`* Corresponding author.
`t Present address: Max-Planck-Institut fiir Biochemie, 8033 Mar(cid:173)
`tinsried, Federal Republic of Germany.
`
`cells made resistant to macrophages display enhanced tu(cid:173)
`morigenicity. Tumor necrosis factor alpha (TN F-a) has been
`shown to play a role in activated macrophage-mediated
`tumor cell killing in vitro (3, 11, 23, 29, 39). NIH 3T3 cells
`transformed by a transfected and amplified HER2/c-erbB-2
`eDNA show increased resistance to the cytotoxic effects of
`activated macrophages or TNF-a in direct correlation with
`increased levels of p185HER2 expression. Furthermore,
`breast tumor cell lines with high levels of p185HER2 exhibit
`resistance to TNF-a. Resistance to host antitumor defenses
`could facilitate the escape of cells from a primary tumor to
`establish metastases at distant sites.
`To further investigate the consequences of alteration in
`HER2/c-erbB-2 gene expression in mammary gland neopla(cid:173)
`sia and to facilitate investigation of the normal biological role
`of the HER2/c-erbB-2 gene product, we have prepared
`monoclonal antibodies against the extracellular domain of
`p185HER2. One monoclonal antibody (4D5) was character(cid:173)
`ized in more detail and was shown to inhibit in vitro
`proliferation of human breast tumor cells overexpressing
`p185HER2 and, furthermore, to increase the sensitivity of
`these cells to the cytotoxic effects of TNF-a.
`
`MATERIALS AND METHODS
`CeUs and cell culture. Human tumor cell lines were ob(cid:173)
`tained from the American Type Culture Collection. The
`mouse fibroblast line NIH 3T3/HER2-3400, expressing an
`amplified HER2/c-erbB-2 eDNA under simian virus 40 early
`promoter control, and the vector-transfected control cell line
`NIH 3T3/CVN have been described previously (18).