`
`##3##*#**##*##*##* 3-81611 292
`ll48 sc1 79‘~
`nazl ”a2
`
`NAIL L10 0
`5
`INDEX
`accc kCCKV
`EEIhesun.
`
`IOEIS X. 143
`
`Page 1
`
`BIOEPIS EX. 1043
`Page 1
`
`

`

`ISSN 0036-8075
`
`6 December 1985 SCIENCE
`
`Volume 230, No. 4730
`
`This Week in Science ......... . ... .. . .. . . . . . ... . .... . .. · · · · · · · · · · · · · · · · · · · ·
`
`LETTERS
`
`Paleontology Renai ssance: C. W. Thayer and C. E. Brett ; Animal Welfare
`Legislation : G. E. Brown, Jr .; Acid Rain Testing: R. H . Es tabrook; NBS
`Budget: D. McClain .... .. ... .... .. ................. .. .. . · ·· · · · · ··· ·· · ··
`
`EDITORIAL
`
`Alzheimer's Disease: A Biologist's Perspectives: C. E. Finch . . . . .... ... · · · · · · · ·
`
`ARTICLES
`
`Human Intelligence: The Model Is the Message: R . J. Sternberg .... ..... · · · · · · ·
`The International Decline in Household Oil Use: L. Schipper and A. N . Keto./f · · ·
`
`Enhanced Transcription of c-myc in Bursal Lymphoma Cells Requires Continuous
`Protein Synthesis: M. Linial, N. Gunderson, M. Groudine ..... . ...... · · · · · ·
`
`Tyrosine Kinase Receptor with Extensive Homology to EGF Receptor Shares
`Chromosomal Location with neu Oncogene: L. Coussens et al.
`. ...... · ·
`
`1091
`
`1106
`
`1109
`
`1111
`
`1118
`
`1126
`
`1132
`
`NEWS AND COMMENT
`
`Politics and Science Clash on African AIDS
`
`Africa and the Origin of AIDS ... . . . .. . ........... . ........ .. .. . .. .. · · · · · · · · ·
`
`Summit Ends with Exchange Agreements .......... ... . ....... .. ............ · ·
`
`Britan Increases Science Spending . . ... ........................... . . . .. · · · · · ·
`
`Pork Barrel Issues Simmer .. ...... . ... . ........ ..... . ........... · · · · · · · · · · · ·
`
`Briefing: U.K. Announces Details of National Space Agency;
`Smithsonian to Feature Information Revolution; USDA Bows to
`Rifkin Call for Review of Seed Bank ; Education: Beginnings of
`Ja~an-U .S . Cooperation; Senate Okays Nuclear Trade Pact with
`Chma ...... ... . .... . . . ......... ... ..... . ..... . · · · · · · · · · · · · · · · · · · · · · · · ·
`
`1140
`
`1141
`
`1142
`
`1144
`
`1145
`
`1146
`
`BOARD OF DIRECTORS
`fCHA.iini~AlNND .
`SECRETARIIS OF
`AAAS SECTIONS
`
`DAVID A. HAMBURG
`Retiring President, Chairman
`
`GERARD PIEL
`President
`
`MATI'IEMATICS (A)
`Daniel Zelinsky
`Lynn Arthur Steen
`
`LAWRENCE BOGORAD
`Presldent·Eiect
`
`~g~~~f ~~~E~~~~ESR
`CHEMISTRY (C)
`Rustum Roy
`Jean'ne M. Shreeve
`
`~~~~Crf N~.~~~~~~~~N'BifR~
`ASTRONOMY (D)
`David Morrison
`John E. Gaustad
`
`PSYCHOLOGY (J)
`John I. Lscey
`William N. Oember
`
`SOCIAL, ECONOMIC, AND POLITICAL SCIE:NCES (K)
`David Mechanic
`David L. Sills
`
`HISTORY AND PHILOSOPHY OF SCIENCE (L)
`Edward Grant
`Arthur L. Norberg
`
`ENGINEERING (M)
`Henry McGee
`W. Edward Lear
`
`EDUCATION (Q)
`John F. Schaff
`Joseph b. Novak
`
`DENTISTRY (R)
`Gordon H. Rovelstad
`Harold M. Fullmer
`
`PHARMACEUTICAL SCIENCES (S)
`Edward G. Ripple
`Belty·ann Hoener
`
`INFORMATION, COMPUTING, AND COMMUNICATION
`Karen B. Levitan
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`
`ARCTIC DIVISION
`
`CARIBBEAN DIVISION
`
`Gunter E. Weller
`Executive Secretary
`
`Juan A. Bonnet, Jr.
`President
`
`BIOEPIS EX. 1043
`Page 2
`
`

`

`AMERICAN ASSOCIATION FOR THE ADVANCEMENT OF SCIENCE
`
`RESEARCH NEWS
`
`Plant Gene Transfer Becomes a Fertile Field. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`Neptune's Ring Arcs Confirmed . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`
`Genes and Biological Clocks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`
`Down Syndrome-Aizheimer's Linked . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`
`1148
`1150
`
`1151
`
`1152
`
`BOOK REVIEWS
`
`... The Heavens and t~e Earth,. reviewed by R. Griffith; Polycyclic
`Hydrocarbons and Carcmo~enesis , S. Broyde; Evolution of Prokaryotes,
`]. A. Shapiro; Books Received ........ . ............................ . ... . .. . .
`
`1154
`
`REPORTS
`
`A Transgenic Mous~ M<;Jdel of the Chronic Hepatitis B Surface Antigen Carrier
`State: F. V. Chtsan et al.
`. ... . . .......... . ... . .. ..... . . .. . . ......... . . .
`
`Specific Expression of Hepatitis B Surface Antigen (HBsAg) in Transgenic Mice:
`C. Babinet, H. Farza , D. Morello, M . Hadchouel, C. Pow·cel . . . . . . . . . . . . . .
`
`1157
`
`1160
`
`Fractal Surfaces of Proteins: M. Lewis and D. C. Rees.. . . . . . . . . . . . . . . . . . . . . . . .
`
`1163
`
`Synthesis and Evaluation of a Prototypal Artificial Red Cell:
`C.A . Huntetal. · ···· · · · · ·· · ·· · ··· ·· · · ········· ·· ·· ·· · · · · · · ··· · ··· ·· · ·
`Biosynthesis and Secretion of Proatrial Natriuretic Factor by Cultured Rat
`Cardiocytes: K. D. Bloch et al.
`. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`
`Human Recombinant Granu.lo~yte-Macrophage Colony-Stimulating Factor: A
`Multilineage Hematop01etm: C. A. Sieff et al.
`. . . . . . . . . . . . . . . . . . . . . . . . . . . .
`
`Identification of a Transcriptional Enhancer Element Upstream from the Proto-
`Oncogene/as: 1. Deschamps, F. Meijlink, I. M. Verma........... .. ...... .
`
`Cloning of a Gene ~hose ~xpression Is Increased in Scrapie and in Senile Plaques
`in Human Bram: S. Wzetgrefe et al.
`. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
`
`Plasticity of Hippocampal Circuitry in Alzheimer's Disease:
`]. W. Geddes et al. · · · · · · · · ·.. .. ....... . ... . .. .. ....... . . ... ...... .... .
`
`1165
`
`1168
`
`1171
`
`1174
`
`1177
`
`1179
`
`COVER
`
`Normal red blood cell and smaller he(cid:173)
`moglobin-carrying artificial red cells
`(ne<?hemocytes) set against an isolated
`capillarJ:'. In the capillary , a 25 percent
`suspension of neohemocytes has re(cid:173)
`placed the blood. The outer membrane
`of the neohemocytes , shown enlarged
`and . cutaway, is a bilayer composed of
`a mixture of four lipids . See page 1165
`[Drawing of Robert Burnett, Chartmas~
`ters, Inc., San Francisco California
`94 133]
`'
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`
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`
`BIOEPIS EX. 1043
`Page 3
`
`

`

`6 December 1985, Volume 230, Number 4730 SCIENCE
`
`AMERICAN ASSOCIATION FOR
`THE ADVANCEMENT OF SCIENCE
`Science serves its readers as a forum for the presentation and
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`Alzheimer's Disease: A Biologist's Perspectives
`
`Public concerns about Alzheimer's disease are rising. With the increasing
`survival to advanced ages, it is predicted that Alzheimer's disease will afflict
`about 2 million people in the United States by the year 2000. Funding for
`basic and clinical studies on this disease has been increased and now
`includes $9 million a year that Congress added to the budget of the National
`Institute on Aging for ten Alzheimer's disease research centers, about $40 -
`million from other National Institutes of Health programs, and $2 inillion
`from private foundations. Biologists may ask how the emphasis on Alz(cid:173)
`heimer's disease could influence support and opportunities for basic research.
`My view is that little recognized but implicit aspects of these programs
`will greatly benefit the neurosciences and biogerontology. An important
`resource will be the greater availability of brain tissues from normal
`subjects. To delineate Alzheimer's disease from other common age-related
`changes requires at least as many (probably several times more) normal
`controls as individuals with Alzheimer's and other age-related dementias.
`Postmortem specimens from normal individuals with detailed personal and
`medical histories are usually scarce. However, healthier relatives and
`friends of victims of Alzheimer's disease are often willing to donate their
`own tissues. The Alzheimer's disease research centers could provide the
`complex logistical support for the short postmortem intervals (4 hours or
`less) needed to preserve many macromolecules and microscopic structures.
`The tissue resources will permit new approaches concerning the impact of
`heredity and environment on the cellular structure and chemistry of the
`healthy human brain. The correlation of detailed pre- and postmortem data
`promises to support major growth of research on human neurobiology and
`could reveal long-lasting effects of drugs, diet, stress, or even subtler
`experiences. Pursuit of these far-reaching and difficult questions will also
`build on the spectacular advances from brain imaging in vivo. Other topics
`so far studied much less in humans than in animals include mechanisms of
`nonischemic neuronal death; cytoskeletal organization; sex differences;
`recep~ors; membrane transport; tissue factors that influence neurite out(cid:173)
`growth; and messenger RNA. The brain messenger RNA's examined at my
`laboratory and that of M. Morrison have a remarkable postmortem stability;
`this invites aggressive use of molecular genetic technology.
`Screening for hereditary influences on Alzheimer's disease could also
`reveal genetic markers linked to depression and other common late-onset
`neurological disorders. Moreover, even without knowing the base sequence
`of an Alzheimer's locus, linked genetic markers could reveal environmental
`factors as well as other genes that influence the age of onset and progress of
`neurological diseases iri high-risk individuals.
`Studies on Alzheimer's disease also probe basic mechanisms of synapto(cid:173)
`genesis. Recently, evidence of neuronal plasticity and sprouting in the
`human brain was found in the hippocampus of victims of Alzheimer's; these
`synaptic reorganizations are similar to the changes induced in the rat
`hippocampus by lesions of the entorhinal cortex.* Intriguing results are
`being obtained by C. Cotman, F. Gage, D. Gash, and others in the use of
`embryonic cell transplants to correct experimental or congenital brain
`lesions that may yield therapies for victims of Alzheimer's. Moreover,
`research leading to the prevention or effective treatment of Alzheimer's
`disease seems likely to illuminate one of the great mysteries in biology-the
`nature of memory and cognition. I would be surprised if the major new
`resources required for a serious attack on Alzheimer's do not also benefit
`the basic neurosciences on the same scale as funding for cancer research
`has done for many areas of molecular, cell, and developmental biology.(cid:173)
`CALEB E. FINCH, Andrus Gerontology Center, Department of Biological
`Sciences, and Alzheimer Disease Research Center Consortium of Southern
`California, University of Southern California, Los Angeles 90089
`
`*J. W. Geddes, D. T. Monaghan, C. W. Cotman, I. T. Lott, R. C. Kim, H. C. Chui, Science, this
`issue, page 1179.
`
`BIOEPIS EX. 1043
`Page 4
`
`

`

`3000 CilmM) (UTP) was then added, and the
`nuclear suspension was incubated at 3o•c for 30
`minutes, after which time 15 ~I ofDNase I (5 ~g/
`ml) in 10 mM CaCI2 (5 ~g/ml) was added. After 5
`minutes at 3o•c, the reaction was made I x SET
`(I percent sodium dodecyl sulfate (SDS), 5 mM
`EDTA, 10 mM tris-HCI, pH 7.4), and proteinase
`K was added to a concentration of 200 ~g/ml.
`After incubation at 37•c fo r 45 minutes, the
`solution was extracted with an equal volume of a
`mixture of phenol and chloroform , and the inter(cid:173)
`phase was again extracted with 100 ~I of 1 x
`SET. Ammonium acetate (I OM) was added to
`ihe combined aqueous phases (original plus
`reextraction) to a final concentration of2.3M, an
`equal volume of isopropyl alcohol was added,
`and nucleic acid '¥a·s precipitated ( -7o·c for 15
`minutes). The precipitate was centrifuged in a
`microcentrifuge for 10 minutes, and the pellet
`was resuspended in 100 ~I of TE (I 0 mM tris(cid:173)
`l:ICI, 1 mM EDTA) and centrifuged through a G-
`50 (medium) spin column. The eluate was made
`0.2M in NaOH and after 10 minutes . on ice,
`HEPES was added to a concentration of 0.24M.
`Two and one-half volumes of ethanol were then
`added, and the soluti on containing the precipi-
`
`tate held overnight at -2o•c. After centrifuga(cid:173)
`tion in a microcentrifuge for 5 minutes , the pellet
`was resuspended in hybridization buffer, which
`consisted of [10 mM TES, pH 7.4, 0.2 percent
`SDS, 10 mM EDTA, 0.3M NaCI , 1 x Den(cid:173)
`hardt's, and Esclrericlria coli RNA (250 ~g/ml)] .
`Nitrocellu lose filters containing plasmid DNA' s
`were prepared with a Schleicher & Schuell Slot
`Blot Apparatus under conditions suggested by S
`and S, except that wells were washed with lO x
`SSC (saline sodium citrate). These filters were
`first hybridized in the hybridization solution
`described above for a minimum of 2 hours at
`65•c. After thi s preliminary hybridization , the
`filters were hybridized to the runoff products in
`hybridization solution for 36 hours. A typical
`reaction contained 2 ml of hybridizat ion solution
`with 1 x 107 cpm/ml. After hybridization , filters
`were washed for 1 hour in 2 X SSC at 65°C. The
`filters were then incubated at 37°C in 2X SSC
`with RNase A (10 mg/ml) for 30 minutes and
`were subsequently washed in 2 x sse at 37•c
`for I hour. Alternatively , after hybridization the
`filters were washed twice for 15 minutes in 0.1
`percent SDS, 2x SSC at room temperature, and
`then washed at 6o•c (0. 1 percent SDS, 0.1 x
`
`SSC) for 30 minutes. Either protocol for proc(cid:173)
`essing of the filters after hybridizatiot1 yielded
`the same specificit y in signal. Filters were then
`exposed to Kodak XAR film in cassettes con(cid:173)
`taining Lightening-Plu s sc reen s at - 7o•c for
`various times.
`45. C. Yanisch-Perron , J . Vierra, J . Me ss ing , Gene
`33, 103 (1985).
`46. S. L. McKnight, E. R. Gavis, R. Kingsbury , R.
`Axel, Cell 25 , 385 (1981) .
`47. M. Groudine and C. Cas imir, Nucleic Acids
`R es. 12, 1427 (1984).
`48. We thank man y of ou r colleagu es for disc ussion
`and suggestions during the course of thi s work;
`Hal Weintraub, Paul Neima n, and Craig Thoml?(cid:173)
`son fo r co mment s on th e manu sc ript ; Cra1g
`Thompson for ass istance in obtaining lympho(cid:173)
`cyte preparat ion s; Bill Schubach for plas mid
`pBK25; and Kay Shiozak i for assistance with
`the manu script. Supported by N IH grants CA
`18282 (M .L.) and CA 28 15 1 (M.L. and M.G .),
`and NSF grant PCM 82-04696 (M.G.), and a
`the Le uk emia Society of
`scholarship from
`America (M.G.)
`
`30 July 1985; accepted 15 Octobe r 1985
`
`RESEARCH ARTICLE
`
`Tyrosine Kinase Receptor with Extensive
`Homology to EGF Receptor Shares
`Chromosomal Location with neu Oncogene
`
`Lisa Coussens, teresa L. Yang-Feng, Yu-Cheng Liao
`Ellson Chen, Alane Gray, John McGrath, Peter H. See burg
`Towia A. Libermann, Joseph Schlessinger, Uta Francke
`Arthur Levinson, Axel Ullrich
`
`In contrast to v-erbB, which encodes a
`68,000-dalton truncated EGF receptor,
`the neu oncogene product is a 185,000-
`dalton cell surface antigen that can be
`detected by cross-reaction with polyclo(cid:173)
`nal antibodies against EGF receptor (I I);
`neu may itself be a structurally altered
`cell surface receptor with homology to
`the EGF receptor and binding specificity
`for an unidentified ligand.
`Using v-erbB as a screening probe, we
`isolated genomic and cDN A clones cod(cid:173)
`ing for an EGF receptor-related , but
`distinct, 138,000-dalton polypeptide hav(cid:173)
`ing all the structural features of a cell
`surface receptor molecu le. On the basis
`of its structural homology , this putative
`receptor is a new member of the tyro(cid:173)
`sine-specific protein kinase family . It is
`encoded by a 4.8-kb messenger RNA
`(tnRNA) that is widely expressed in nor(cid:173)
`mal and malignant ti ss ue s. We have lo(cid:173)
`calized the gene for this protein to q21 of
`chromosome 17, which is di stinct from
`the EGF receptor locus , but coincident
`with the neu oncogene mapping position
`(12). We therefore consider the possibili(cid:173)
`ty that we have isolated and character(cid:173)
`ized the normal human counterpart of
`the rat neu oncogene.
`Tyrosine kinase-type receptor gene and
`complementary DNA. As part of our at(cid:173)
`tempts to isolate and characterize the
`chromosomal gene coding for the human
`cellular homologue of the viral erbB gp68
`polypeptide, AEV-ES4 erbB sequences
`(2.5-kb Pvu II fragment of pAEV) (13)
`were used as a 32P-Iabeled hybridization
`probe for the screening of a human geno(cid:173)
`mic DNA library at reduced stringency
`
`lin (6), PDGF (7), and insulin-like growth
`factor 1 (IGF- 1) (8); hence more connec(cid:173)
`tions may be found between tyrosine
`kinase growth factor receptors and tyro(cid:173)
`sine kinase oncogene products.
`Comparison of the complete primary
`structure of the human EGF receptor (9)
`with the sequence of the av ian erythro(cid:173)
`blastosis virus (AEV) transforming gene,
`v-erbB (iO), revealed close seq uence
`similarity ; in addition, there were amino
`and carboxyl terminal deletions that may
`reflect key structural changes in the ge n(cid:173)
`eration of an oncogene from the gene for
`a normal growth factor receptor (3, 9).
`Another oncogene, termed neu, is also
`related to v-erbB and was originally
`identified by its activation in ethylnitro(cid:173)
`sourea-i nduced rat neuroblastomas (II).
`
`Growth factors and their receptors are
`involved in the regulation of cell prolif(cid:173)
`eration , and several recent findings sug(cid:173)
`gest that they _ also play a key role in
`oncogenesis (1-4). Of approximately 20
`identified oncogenes, th.e three that have
`been correlated with known cellular pro(cid:173)
`teins are each related to either a growth
`factor or a growth factor receptor. The B
`chain of platelet-derived growth factor
`{PDGF) is encoded by the proto-onco(cid:173)
`gene c-sis (2), the erb-B oncogene prod(cid:173)
`uct gp68 is a truncated form of the epi(cid:173)
`dermal growth factor (EGF) receptor (3),
`and the proto-oncogene c-fms may be
`related or identical to the receptor for
`macrophage colony-stimulating factor
`(CSF-lR) (4).
`The receptor-related oncogenes are
`members of a gene family in that each
`has tyrosine-specific protein kinase ac(cid:173)
`tivity, and is associated with the plasma
`membrane (5) . Such features are also
`shared by several other polypeptide hor(cid:173)
`mone receptors, including those for insu-
`
`11 32
`
`Lisa Coussens, Yu-Cheng Liao, Elison Chen, Alane Gray , Peter H . Seeburg, Arthur Levinson , and Axel
`Ullrich are in the Department of Molecular Biology , Genentech , Inc., 460 Point San Bruno Boulevard, South
`San Francisco, California 94080; John McGrath is currently with the Department of Biology , Massac hu setts
`Institute of Technology , Cambridge, Massachusetts 02142; Towia Libermann ahd Jose ph Schlessinger are in
`the Department of Chemical Immunology at the Weizmann Institute of Science, Rehovot 76100, Israe l; and
`Teresa L. Yang-Feng and Uta Francke are in the Department of Human Genetics at Yale University Sc hool
`of Medicine, 333 Cedar Street , New Haven, Connecticut 065 10.
`
`SCIENCE, VOL. 230
`
`BIOEPIS EX. 1043
`Page 5
`
`

`

`(14 ). Clone A.c-erbB/ 1 was isolated ; it
`contained a hybridizing 1. 8-kb Bam HI
`fragment, whi ch was subjected to DNA
`sequence analysis. T he 1838-bp se(cid:173)
`quence contains three complete and one
`partial erbB-homologous exons separat(cid:173)
`ed by short intervening sequences (Fig.
`1) . Co mparison of thi s human gene se(cid:173)
`quence with our compl ete eDNA-de(cid:173)
`ri ved human EGF receptor protein se(cid:173)
`quence (9) revealed 32 di{ferences (1 8. 7
`perce nt) within
`the 171 amino acid
`stretch of combined exons, suggesting
`that this gene fragment was not deri ved
`from th e human EGF receptor gene.
`Since thi s gene may code for a n un(cid:173)
`tyrosine kinase-type receptor
`known
`that is closely related to th e human EGF
`receptor, we named it H ER2 .
`Northern blot analysis (/ 5) with the
`32P-Iabeled 1.8-kb H ER2 fragment as a
`hybridi zation probe revealed a 4.8-kb
`mRN A in human term placenta pol y(At
`RNA , di stinct from the 5.8- and 10.5-kb
`EGF rece ptor mRN A' s also present at
`hi gh levels in this ti ssue (f ig. 2a , lane 1).
`Thus, we had isolated a portion of an
`EG F rece ptor- erbB-related but di stinct
`gene. To obtain its complete primary
`structure, two single-stranded sy nthetic
`oligonucl eotide probes (16) were pre(cid:173)
`pared from HER2 exon sequence regions
`th at differed sufficientl y (less than 60
`percent nu cleotide sequence homology)
`from EGF receptor DNA sequences
`(Fig. I, I and 2) and used to screen
`a term placenta compl ementary DNA
`(e DNA) library of 2 x 106 independent
`recombinant clo nes in A.gtiO (1 7). Fifty(cid:173)
`two clones were isolated ; they hybrid(cid:173)
`ized strongly with both synthetic probes
`and weakl y with an EG F receptor eDNA
`fragment (H ER64-3) (9) containing the
`homologous region within the tyrosine
`kinase domain . One of these , t..H ER2-
`436, had the longest e DNA inse rt (4.5
`kb), consisting of three Eco Rl fragments
`(1.4 , 1.5 , and 1.6 kb).
`T he compl ete eDN A sequence of this
`clone is shown in Fig. 3. T he longest
`open reading fra me starting with a me(cid:173)
`thi onine codon codes for a 1255 amino
`acid polypeptide ( 137 ,828 daltons) and
`contains the 171 residues encoded by the
`four exons in th e 1.8-kp HER2 gene Bam
`HI fragment (Fig. 1). Thi s 3765-bp cod(cid:173)
`ing sequence is fl anked by 150 bp of 5'
`untranslated sequence and a TGA stop
`codon, fo llowed by a 627-nucleotid e 3'
`untranslated sequ ence . No stop codon is
`found in the 5' untranslated region. In
`support of our assignment, however , the
`initiati on codon at position 151 is fl anked
`by sequences that follow perfect ly Ko(cid:173)
`za k' s rul e (/8) for translation initiation.
`The 3' untranslated sequence contains a
`
`6 DECEMBER 1985
`
`potential poly(A) addition signal se(cid:173)
`quence (AAT AT A) 12 nucl eotides up(cid:173)
`stream from a stretch of 15 adenylate
`residues. We are not certain if this (A) 15
`stretch is part of a poly(A) tail or repre(cid:173)
`sents an internal pol y(A) stretch of a
`longer 3' untranslated sequence.
`
`those for EGF and insulin (9, 19). Such
`features are apparent in the hydropathy
`profile (20) compari son (Fig. 4a). On the
`basis of thi s comparison , and on amino
`acid sequence alignment with the EGF
`receptor (Fig. 4b , region 1), we predict a
`21 amino acid signal sequence (Fig. 4b,
`
`Abstract. A novel potential cell surface receptor of the tyrosine kinase gene family
`has been identified and characterized by molecular cloning. Its primary sequence is
`very similar to that qf the human epidermal growth f actor receptor and the v-erbB
`oncogene produ ct; the chromosomal location of
`the gene for this protein is
`coincident with the neu oncogene , which suggests that the two genes may be
`identical.
`
`Comparison of EGF receptor and
`HER2 sequence. As already indicated by
`the v-erbB sequence homology used to
`isolate HER2 , the putative HER2 pro(cid:173)
`tein is very similar in its overall domain
`organization and sequence to the EGF
`receptor. Nevertheless , there are differ(cid:173)
`ences that are likely to define a specific
`biological role for the HER2 polypep(cid:173)
`tide.
`The predicted HER2 polypeptide con(cid:173)
`tains each of the domain features found
`in hormone receptor precursors, such as
`
`1) , an amino terminal serine residue , and
`a 632 amino acid putative extracellular
`ligand-binding domain; a highly hydro(cid:173)
`phobic , 22-amino acid transmembrane
`anchor domain separates the extracellu(cid:173)
`lar domain from a 580-residue-long car(cid:173)
`boxyl-terminal
`cytoplasmic
`domain ,
`which possesses the highest homology to
`v-erbB and other members of the tyro(cid:173)
`sine kinase fa mily.
`The 632-amino acid , putative HER2
`ligand binding domain is about 40 per(cid:173)
`cent homologous with the 621-residue
`
`769
`Al a
`Glu
`Lys
`740 Gl u
`11 cProAs pG 1 yG l uA snVa ll ys l l cPr oVa 1 A 1 a 11 elys Va 1 LeuArgG 1 uAsn ThrSe rPr olysA 1 aAs nl ysG 1 u l l el cuAs p
`1 GGA T CCC TGA TGGGGAGM TGT GMM TTCCAG TGGCCA TCMAGTGTTGAGGGAAAACACA TCCCCCMAGCCAACAMGMATCT T AGACGT AAGCCCCTCCACCCTCT CCI GCTAGG
`
`111 AGGACAGGMGGAC CCCA TGGC TGCAGG T C T GGGCT C T GG T CT C T C TT CA TT GGGG TIT GGGGAGA TAT GAC T CC CGCAMC C T AGAC T A TT TT TT T GGAGAC GGAGCT TGC T C T G T CAC
`
`14 1 CC AGGCT GGAG T GCAG TGGC G TT AT C T CGGC T CAC TGCMC CT CCACC T C C T GGAC T CAAGCGA TT T T CA T GC CT CAGGC T CC T GAG T AGC TGGGA TT ACAAGC GC C CGC T AA T T T T TT T
`
`36 1 T TTTTT T TT GAGACAGAG T CTCGC T C T G T CAC CCAGGCT AGAG T GMA T GG TGCGG T CT CAGCT CAGC CT C CCAGG T T MAGCGA T T C TT C T CCC T CAG T C T CCTGAG T AGCT GGGA T TA
`
`481 CAGGCGCGAGCCACCACGCCCGGCT M TTT TTGTAT T ITTAGT A GAGA TGGGA TT TCACCATG ITGGCCAGGTTGGTGTCAAACTCC TGACCTCA TGATCCGCCCGCCTCGGCCTCCCAA
`
`60 I AG T GC T GGGA T T ACAGG TG T GAGCCACG T GCCCGGCCT M T CIT T G T A TT T T TAG TAGAGACAGGG TTT CACCA TG TT G T CCAGGC TGG TAC T T TGAGCCT T CACAGGCTG TGGGCCA TG
`
`Hf
`AspA sn
`Ser
`770
`G 1 uA 1 aTyr Va !Met A 1 aG l yVa l Gl ySer Pr oTy
`71 1 GCTGTGGT TTGTGA TGGTTGGGAGGCTGTGTGGTGTITGGGGGTGTGT GGTCTCCCA T ACCC TCTCAGCGT ACCC T TGTCCCCAGGAAGCA TACGTGA TGGCTGGT GTGGGCTCCCCATA
`
`Ty
`His lysAs pA snll e
`Tyr
`Phe
`li e
`Cys
`s
`rVa l SerArgleulc uG ly l 1 eCysLeuTh rSc r ThrV a 1 G 1 nl euV a I ThrG l nle uMetPr oTyr G l yCysle uleuAspll i s Va l Ar gG l uAsnAr gG lyArgl euG l ySe rGl nAs
`84 1 TG T C T C C CGC C TT CT GGGCA T CT GCC T GACA T C CACGG T GCAGCTGG T GAC AC AGC IT AT G CCC TAT GG CT GCC T C IT AGAC CA T G T C CGGGA AAAC CG CGGACGC C T GGG C T C CCAGGA
`
`83 1
`Val
`r
`pl e ulcuAs n Trp Cys~le lG 1 n I leA 1 alys
`96 1 CC TGCTGMC TGGTGT ATGCAGA TTGCCMGGTA TGCACC TGGGC TCITTGCAGG TCTCT CCGGAGCAAACCCC TATGTCCACAAGGGGC T AGGATGGGGACTC TTGC TGGGCA TGTGGC
`
`Thr
`Arg
`Asn
`832
`G 1 yMetSer Tyrl euG 1 uAspVa 1 Ar qleuVa 1 Hi sAr gAspleuA 1 aA 1 aAr gAsnVa ll euV a 1 LysSer
`I 081 CAGGC C CAGGCCC T C C CAGMGGT C T ACA T GGG TGC T T CC CAT T C CAGGGGA TGAGC T AC CT GGAGGA T G T GCGG CT CG TACACAGGGAC T T GGCCGCT CGGMCG TGC T GG T CAAGAGT
`1-CCGT- CT-G--G - C-C---C-
`- A- I(D
`
`883
`Gl u
`Gl yA l aG l u
`Lys
`Gi n
`l ys
`Pr o;\snfl i s Va l l ys II elhrAspPheG lyleuA I aArgLeul e uAsp l leAspG 1 uThr G l uTyr iH s A 1 aAs pG lyG lyl ys
`1101 CCCAACCATGTCAAM TTACAGACITCGGGC TGGCTCGGCTGCTGGACAT TGACGAGACAGAG TACCATGCAGATGGGGGCAAGGT T AGGTGMGGACCAAGGAGCAGAGGAGGCTGGG T
`1- CMA-- -----GTGCG A- 1@
`
`884
`Va 1 Pr o I le l ysTrpMet A I aleuG 1 uSer I 1 el eu
`1 3 21 GGAG TGG T G T C T AGC CC AT GGGAGAAC T C T GAG l GGCCACT CC CACM CA CACAG TT GGAGGAC TT CC T CTT CTGC CC T CCC CAG T GCC CAT CAAG T GGA T GGCGC TGGAGT C CA TT CT C
`
`910
`Il eTyr
`Hi s
`ArgA rgAr gPhe ThrH i sG 1 nSer As pVa 1 TrpSerTyr Gl yVa 1
`144 1 CGCCGGCGG TTCACCCACCAGAGTGATGTGTGGAGTT ATGG TGTGTGATGGGGGG TG ITGGGAGGGGTGGGTGAGGAGCCA TGGC TGGAGGGAGGA TGAGAGCTGGGA TGGGGAGAA TT A
`
`15 61 C GGGGC CACC T CAGCA T G TGAAGGGAGGGMGGGGC T GCCT G T GCC CCAC C T T G CAGGG T CT G T GCAC TT C C CAGGA T T AGGGMAGAC CGG G T AGGG T C T G T C T C CT GGC AT CA CAT CT
`
`1681 CCC C C T GCT AC CT GCCA T GA T GCT AGAC T C CT GAGC AGAACC T C T GGC T CAG T ACACT AAAGCT C CCT CT GGC CCT CCC ACT CC T GAC C CTGT C T C T GC CTT AGG TG T GAC T G T G T GGGA
`
`1 BO I GC TGA TGAC TTTT GGGGCCAMCC TT ACGATGGGATCC
`
`Fig. I . Partial sequence of the H ER2 gene. A partial Hae rii-Aiu I genomic library (14 ) of human
`fe tal DNA in >-. Charon 4A was screened using a radiolabeled 2.5-kb Pvu II fragment of pAEV
`(13) containing coding sequences fo r the tyrosine kinase domain . Hybridization was as
`described el