September 3, 2013
`
`
`vol.110 I no. 35 I pp. 1450544814 '-
`
`'I'roceedings of the National Academy of Sciences of the United States of America
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`

`

`Cover image: Pictured is an aggregate of dolomite nanocrystals on the surface of a carboxylated
`polystyrene sphere. The mineral dolomite is abundant in the geologic record and an be synthesized
`in the laboratory at high temperatures, but the mechanism of its formation at low temperatures
`remains unclear. Jennifer A. Roberts et a]. demonstrate that mrboxy] groups on the surfaces of
`polystyrene spheres can catalyze dolomite precipitation at 30°C from solutions that simulate
`seawater. The findings suggest that natural surfaces that possess a high density of carboxyl groups,
`such as microbial biomass, may be key to the formation of dolomite in low temperature sedimentary
`systems. See the article by Roberts et al. on pages 14540-14545. Image courtesy of David S. Moore.
`
`From the Cover
`
`14610 Biological role of a polyglutamine tract
`
`14540 Dolomite formation at low temperature
`
`14520 Neutralization of differentiation in gliomagenesis
`
`14528 Characterizing a bacteria-amoeba symbiosis
`
`14546 Modeling ancient atmospheric oxygen levels
`
`1450? Spinning silk from spiders
`Charles Choi
`
`
` 14505
`In This Issue
`14508 QnAs with Ronald DePinho
`Prasltant Nair
`-O See Inaugural Article on page 14520
`
`14510
`
`E3368 Lack of chronological support for stepwise prehurnan
`extinctions of Australian megafauna
`Barry W. Brook, Corey l. A. Bradshaw, Alan Cooper,
`Christopher N. Johnson. Trevor H. Worthy, Michael Bird,
`Richard Gillespie, and Richard G. Roberts
`
`Changing the fate of cancer. one splice at a time
`David L. Schonberg, Monica Venere, and Jeremy N. Rich
`4 See companion article on page 14520
`
`E3369 Reply to Brook et al: No empirical evidence for human
`overkill of megafauna in Sahul
`Stephen Wroe. Judith H. Field, Michael Archer, Donald K.
`Grayson, Gilbert J. Price, Julien Lcuys, J. Tyler Faith.
`Gre o
`E. Webb, Iain Davidson, and Scott D. Moone
`y
`g ry
`E3370 Concerns over interspecies transcriptional comparisons
`in mice and humans after trauma
`Andrew R. Osterburg, Philip Hexley, Dorothy M. Supp,
`Chad T. Robinson, Greg Noe], Cora Ogle, Steven T. Boyce,
`Bruce J. Aronow, and George F. Babcock
`
`E3371 Reply to Osterburg et al.: To study human inflammatory
`diseases in humans
`Ronald G. Tompkins, H. Shaw Warren, Michael N.
`Mindrinos, Wenzhong Xiao, and Ronald W. Davis
`
`J
`
`* Free online through the PNAS open access option.
`
`14512 Small molecules mediate bacterial farming by
`social amoebae
`Robert A. Raguso
`.. See companfbn [gm-dc 0,, page {4523
`14514 A cold, hard look at ancient oxygen
`Boswell A. Wing
`-> See companion article on page 14546
`
`14516 A flexible polyglutamine hinge opens new doors for
`understanding huntingtin function
`Flaviano Giorgini
`-) See companion article on page new
`
`
`
`14518 Significance Statements
`-9 Bnefsrarements wn'rren by the authors about the Significance
`of their papers.
`
`PNAS | 5epternber3.2013 | vol.110 | no. 36 |
`
`Iii-vii
`
`

`

`
`
`14592
`
`14520
`
`Neutralization of terminal differentiation
`in gliomagenesis
`Jian Hu, Allen L. Ho, Liang Yuan. Baoli Hu, Sujun Hua,
`Soyoon Sarah Hwang. Jianhua Zhang, Tianyi Hu, Hongwu
`Zheng, Boyi Gan. Gongxiong Wu, Yani Alan Wang.
`Lynda Chin, and Ronald A. DePinho
`-I See QM: on page 14508
`-I See Commentary on page 14510
`
`
`
`CHEMISTRY
`
`E3445
`
`14528
`
`14504
`
`14653
`
`14534
`
`14540
`
`14546
`
`14551
`
`Stapled inc—helical peptide drug development: A potent
`dual inhibitor of MDM2 and MDMX for pSB-dependent
`cancer therapy
`Yong 5. Chang, Bradford Graves, Vincent Guerlavais.
`Christian Tovar, Kathryn Packman, Kwong~Him To, Karen A.
`Olson. Kamala Kesavan, Pranoti Gangurde, Aditi Mukherjee,
`Theresa Baker, Knysztof Darlak. Carl Elkin, Zoran Filipovic,
`Farooq Z. Qureshi. Hongliang Cai, Pamela Berry. Eric
`Feyfant, Xiangguo E. Shi, James Horstick, D. Allen Annis.
`Anthony M. Manning, Nader Fotouhi. Huw Nash,
`Lyubomir T. Vassilev, and Tomi K. Sawyer
`
`A bacterial symbiont is converted from an inedible
`producer of beneficial molecules into food by a single
`mutation in the gacA gene
`Pierre Stallforth, Debra A. Brock, Alexandra M. Cantley.
`Xiangjun Tian, David C. Queller, Joan E. Strassmann.
`and Jon Clarcly
`-D See Commentary on page 14512
`
`Pulsed hydrogen—deuterium exchange mass
`spectrometry probes conformational changes in
`amyloid beta {All} peptide aggregation
`Ying Zhang, Don L. Rempel. Jun Zhang, Anuj K. Sharrna,
`Liviu M. Min'ca. and Michael L. Gross
`
`Axial interactions in the mixed-valent Cu,“ active site and
`role of the axial methionine in electron transfer
`Ming-Li Tsai, Ryan G. Hadt, Nicholas M. Marshall,
`Tiffany D. Wilson, Yi Lu, and Edward I. Solomon
`
`COMPUTER SCIENCES
`
`Efficient discovery of overlapping communities in
`massive networks
`Prem K. Gopalan and David M. Blei
`
`EARTH. ATMOSPHERE. AND PLANETARY SCIENCES
`
`Surface chemistry allows for abiotic precipitation of
`dolomite at low temperature
`Jennifer A. Roberts, Paul A. Kenward, David A.
`Fowle, Robert H. Goldstein, Luis A. Gonzalez,
`and David S. Moore
`
`Dynamic model constraints on oxygen-17 depletion in
`atmospheric 0, after a snowball Earth
`Xiaobin Cao and Huiming Bad
`4 See Commentary on page 14514
`
`Synchronous interhemispheric Holocene climate trends
`in the tropical Andes
`Pratigya .l. Polissar. Mark B. Abbott, Alexander P. Wolfe,
`Mathias Vuille. and Maximiliano Bezada
`
`'rv
`
`I www.pnas.org
`
`ENGINEERING
`
`Design and characterization of synthetic fungal-bacterial
`consortia for direct production of isobutanol from
`cellulosic biomass
`
`Jeremy J. Minty, Marc E. Singer, Scott A. Scholz,
`Chang-Hour: Bae, Jung-Ho Ahn, Clifton E. Foster,
`James C. Liao, and Xiaoxia Nina Lin
`
`STATISTICS
`
`14557
`
`14563
`
`On robust regression with high-dimensional predictors
`Noureddine El Karoui, Derek Bean, Peter J. Bickel,
`Chinghway Lim, and Bin Yu
`
`Optimal nil-estimation in high-dimensional regression
`Derek Bean, Peter J. Bickel, Noureddine El Karoui.
`and Bin Yu
`
`
`
`14569
`
`14574
`
`14580
`
`14586
`
`ANTHROPOLOGY
`
`Environmental productivity predicts migration,
`demographic. and linguistic patterns in
`prehistoric California
`Brian F. Codding and Terry L Jones
`
`PSYCHOLOGICAL AND COGNITIVE SCIENCES
`Neural mechanisms of communicative innovation
`Arjen Stolk, Lennart Verhagen, Jan-Mathijs Schoffelen.
`Robert Oostenveld, Mark Blokpoel, Peter Hagoort,
`Iris van Rooij, and Ivan Toni
`
`Sight over sound in the iudgment of music performance
`Chia-Jung Tsay
`
`Ontogeny of prosocial behavior across diverse societies
`Bailey R. House, Joan B. Silk, Joseph Henrieh, H. Clark
`Barrett. Brooke A. Scelza, Adam H. Boyette, Barry S.
`Hewlett, Richard McElreath, and Stephen Laurence
`
`
`
`14569
`
`14586
`
`14592
`
`ANTHROPOLOGY
`
`Environmental productivity predicts migration.
`demographic. and linguistic patterns in
`prehistoric California
`Brian F. Codding and Terry L. Jones
`
`Ontogeny of prosocial behavior across diverse societies
`Bailey R. House, Joan B. Silk, Joseph Henrich, H. Clark
`Barrett, Brooke A. Scelza, Adam H. Boyette, Barry S.
`Hewlett, Richard MeElreath, and Stephen Laurence
`
`APPLIED BIOLOGICAL SCIENCES
`
`Design and characterization of synthetic fungal-bacterial
`consortia for direct production of isobu'tanol from
`cellulosic biomass
`Jeremy J. Minty, Marc E. Singer. Scott A. Scholz,
`Chang-Hoon Bae, Jung-Ho Ahn, Clifton E. Foster,
`James C. Liao, and Xiaoxia Nina Lin
`
`

`

`14598
`
`14604
`
`14610
`
`14616
`
`14622
`
`14628
`
`14634
`
`14640
`
`14646
`
`14652
`
`E3372
`
`E3381
`
`Engineered knottin peptide enables noninvasive optical
`imaging of intracranial medulloblastoma
`Sarah J. Moore, Melanie G. Hayden Gephart, Jamie M.
`Bergen, YouRong S. Su, Helen Rayburn, Matthew P. Scott,
`and Jennifer R. Cochran
`
`BIOCHEMISTRY
`
`Pulsed hydrogen-deuterium exchange mass
`spectrometry probes conformational changes in
`amyloid beta (Ail) peptide aggregation
`Ying Zhang, Don L. Rempel, Jun Zhang, Anuj K. Sharma,
`Liviu M. Mirica. and Michael L. Gross
`
`Polyglutamine domain flexibility mediates the proximity
`between flanking sequences in huntingtin
`Nicholas Stephane Caron, Carly Robyn Desmond,
`Jianrun Xia, and Ray Truani
`-D See Commentary on page 145M
`
`Glucocorticoid receptor represses proinflammatory
`genes at distinct steps of the transcription cycle
`Rebecca Gupte, Ginger W. Muse, Yurii Chinenov,
`Karen Adelman, and Inez Rogatsky
`
`Analysis of Chlamydomonas thiamin metabolism in vivo
`reveals riboswitch plasticity
`Michael Moulin, Ginnie T. D. T. Nguyen, Mark A. Scaife.
`Alison G. Smith, and Teresa B. Fitzpatrick
`
`Human a—L—iduronidase uses its own N—glycan as
`a substrate-binding and catalytic module
`Nobuo Maita, Takahiro Tsukimura, Takako Taniguchi. Seiji
`Saito, Kazuki Ohno, Hisaaki Taniguchi, and Hitoshi Sakuraba
`
`Insights into Eph receptor tyrosine kinase activation
`from crystal structures of the EphA4 ectodomain and its
`complex with ephrin-AS
`Kai Xu, Dorothea Tzvetkova-Robev, Yan Xu, Yehuda
`Goldgur. Yee—Peng Chan, Juha P. Himanen,
`and Dimitar B. Nikolov
`
`Cotranslational folding of membrane proteins probed by
`arrest-peptide—mediated force measurements
`Florian Cymer and Gunnar von Heijne
`
`Glycosylated linkers in multimodular
`lignocellulose-degrading enzymes dynamically bind
`to cellulose
`
`Christina M. Payne, Michael G. Resch, Liqun Chen,
`Michael F. Crowley, Michael E. Himmel, [any E. Taylor I[.
`Mats Sandgren, Jerry Stfihlberg, Ingeborg Stals, Zhongping
`Tan, and Gregg T. Beckham
`
`Implications for the evolution of eukaryotic
`amino-terminal acetyltransferase (NAT) enzymes
`from the structure of an archaeal ortholog
`Glen Liszczak and Ronen Marmorstein
`
`14658
`
`14664
`
`14670
`
`E3388
`
`14520
`
`14676
`
`Axial interactions in the mixed-valent Cu; active site and
`role of the axial methionine in electron transfer
`Ming—Li Tsai, Ryan G. Hadt, Nicholas M. Marshall.
`Tiffany D. Wilson. Yi Lu, and Edward 1. Soiomon
`
`Structure of the YajR transporter suggests a transport
`mechanism based on the conserved motif A
`Daohua J iang, Yan Zhao, Xianpirig Wang, Junping Fan.
`Jic Heng, Xuehui Liu, Wei Feng, Xusheng Kang, Bo
`Huang, J ianfeng Liu, and Xuejun Cai Zhang
`
`The growth speed of microtubules with
`XMAP215-coated beads coupled to their ends
`is increased by tensile force
`Anastasiya Trushko, Erik Schafier, and Jonathon Howard
`
`CELL BIOLOGY
`
`Transcriptional response to stress in the dynamic
`chromatin environment of cycling and mitotic cells
`Anniina Vihervaara, Christian Sergelius, Jenni Vasara,
`Malin A. H. Blom, Alexandra N. Elsing, Pia Roos-Mattjus,
`and Lea Sisionen
`
`Neutralization of terminal differentiation
`in gliomagenesis
`J iari Hu, Alien L. Ho, Liang Yuan, Baoli Hu. Sujun Hua,
`Soyoon Sarah Hwang, Jianhua Zhang, Tianyi Hu, Hongwu
`Zheng, Boyi Gan, Gongxiong Wu, Yaoqi Alan Wang,
`Lynda Chin, and Ronald A. DePinho
`4 See QnAs on page [4508
`-) See Commentary on page 14510
`
`NAP-1 and mitoNEET are central to human breast cancer
`proliferation by maintaining mitochondrial homeostasis
`and promoting tumor growth
`Yang—Sung Sohn, Sagi Tamir, Luhua Song, Dorit Michaeli,
`Imad Matouk, Andrea R. Conlan, Yacl Harir, Sarah H. Holt,
`Vladimir Shulaev, Mark L. Paddock, Abraham Hochberg,
`loav Z. Cabanchiek, 1036 N. Onuchic, Patricia A. Jennings.
`Rachel Nechushtai, and Ron Mittler
`
`14682
`
`14688
`
`DEVELOPMENTAL BIOLOGY
`
`Alteration of genic 5-hydroxymethylcytosine patterning
`in olfactory neurons correlates with changes in gene
`expression and cell identity
`Bradley M. Colquitt, William E. Allen, Gilad Barnea.
`and Stavros bomvardas
`
`Torso-like functions independently of Torso to regulate
`Drosophil'a growth and developmental timing
`Travis K. Johnson, Tova CrOSSman, Karyn A. Foote,
`Michelle A. Henstridge, Melissa J. Saligari, Lauren
`Forbes Beadle. Anabel Herr, James C. Whisstock,
`and Coral G. Warr
`
`BIOPHYSICS AND COMPUTATIONAL BIOLOGY
`
`14528
`
`Two-state dynamics of the SH3—SH2 tandem of Abl
`kinase and the aliosteric role of the N-cap
`Carles Corbi-Verge, Fabrizio Marinelli. Ana Zafra—Ruano,
`Javier Ruiz-Sana, Irene Luque, and .1056 D. Faraldo—Gomez
`
`Consequences of domain insertion on
`sequence-structure divergence in a superfold
`Chetanya Pandya, Shoshana Brovrn, Ursula Picper, Andrej
`Sali, Debra Dunaway-Mariano, Patricia C. Babbitt. Yu Xia,
`and Karen N. Allen
`
`ECOLOGY
`
`A bacterial symbiont is converted from an inedible
`producer of beneficial molecules into food by a single
`mutation in the gacA gene
`Pierre Stallforth. Debra A. Brock, Alexandra M. Cantley,
`Xiangjun Tian, David C. Queller, Joan E. Strassmann,
`and Jon Ciardy
`-D Sec Commentary on page i 451 2
`
`14693
`
`A physical explanation of the temperature dependence
`of physiological processes mediated by cilia and flagella
`Stuart Humphries
`
`PNAS | September3.2013 | vo|.110 | no.36 | v
`
`

`

`EVOLUTION
`
`14735
`
`14699
`
`E3398
`
`E3408
`
`E3417
`
`14705
`
`14711
`
`E3425
`
`14717
`
`14723
`
`14729
`
`Segmenting the human genome based on states of
`neutral genetic divergence
`Prabhani Kuruppurnullage Don, Guruprasad Ananda,
`Francesca Chiaromonte, and Kateryna D. Makova
`
`GENETICS
`
`Establishing a hematopoietic genetic network through
`locus~specific integration of chromatin regulators
`Andrew W. DeVilbiss, Meghan E. Boyer,
`and Emery H. Bresnick
`
`Inherited mutations in the helicase RTELl cause telomere
`dysfunction and Hoyeraal—Hreidarsson syndrome
`Zhong Deng, Galina Glousker, Allah Molezan,
`Alan J . Fox, Noa Lamm, J ayaraju Dheekollu, Orr~El
`Weizman, Michael Schertzer, Zhuo Wang, Olga
`Vladimirova, Jonathan Schug, Memet Aker, Arturo
`Londono-Vallejo, Klaus H. Kaestner, Paul M. Lieberman,
`and Yehuda Tzfati
`
`Assembler for de novo assembly of large genomes
`Te—Chin Chu, Chen-Hua Lu, Tsunglin Liu, Greg C. Lee,
`Wen—Hsiung Li, and Arthur Chun—Chieh Shih
`
`Gene duplication in the major insecticide target site. Rdl.
`in Drosophila melanogaster
`Emily J. Remnant, Robert T. Good, Joshua M. Schmidt,
`Christopher Lumb, Charles Robin, Phillip J. Daborn,
`and Philip Batterham
`
`.
`
`IMMUNOLOGY
`
`Blockade of Au receptors potently suppresses the
`metastasis of Cl'J'B+ tumors
`Paul A. Beavis, Upulie Divisekera, Christophe
`Paget, Melvyn T. Chow, Liza B. John, Christel
`Devaud, Karen Dwyer, John Stagg. Mark J. Smyth,
`and Phillip K. Darcy
`
`MEDICAL SCIENCES
`
`Hypoxic retinal Miiller cells promote vascular
`permeability by HIM—dependent up-regulation of
`angiopoietin-like 4
`Xiaoban Xin, Murilo Rodrigues, Mahaa Umapathi, Fabiana
`Kashiwabuchi, Tao Ma, Savalan Babapooerarrokhran,
`Shuang Wang, Jiadi Hu, Imran Bhutto, Derek S.
`Welsbie, Elia .l. Duh, James T. Handa, Charles G.
`Eberhart, Gerard Lutty, Gregg L. Semcnza.
`Silvia Montaner, and Akrit Sodhi
`
`A nanoparticle formulation that selectively transfects
`metastatic tumors in mice
`
`J ian Yang, William Hendricks, Guosheng Liu, J. Michael
`McCaffery, Kenneth W. Kinzler, David L. Huso,
`Bert Vogelstein, and Shibin Zhou
`
`Retinoblastoma tumor suppressor protein in pancreatic
`progenitors controls a- and fl-cell fate
`Erica P. Cai, Xiaohong Wu, Stephanie A. Schroer,
`Andrew J. Elia, M. Cristina Nostro, Eldad Zacksenhaus,
`and Minna Woo
`
`PINK1 regulates histone H3 trimethylation and gene
`expression by interaction with the polycomb
`protein EEDIWAIT1
`Arnaud Berthier, Judit Jiménez‘Séinz, and Rafael Pulido
`
`vi
`
`www.pnas.org
`
`14540
`
`14741
`
`14747
`
`14753
`
`E3435
`
`14574
`
`14759
`
`14765
`
`14771
`
`14777
`
`Protein kinase Cl} as a therapeutic target stabilizing
`blood—brain barrier disruption in experimental
`autoimmune encephalomyelitis
`Tobias V. Lanz, Simon Becker, Matthias Osswald, Stefan
`Bittner, Michael K. Schuhmann, Christiane A. Opitz,
`Sadanand Gaikwad, Benedikt Wiestler, Ulrike M.
`Litzenburger, Felix Sahm, Martina Ott, Simeon Iwantscheff,
`Carl Grahitz. Michel Mittelbronn. Andreas von Deimling,
`Frank Winkler, Sven G. Meuth, Wolfgang Wick,
`and Michael Flatten
`
`MICROBIOLOGY
`
`Surface chemistry allows for abiotic precipitation of
`dolomite at low temperature
`Jennifer A. Roberts, Paul A. Kenwarcl, David A.
`Fowle, Robert H. Goldstein, Luis A. Gonzalez,
`and David S. Moore
`
`Single point mutations in ATP synthase compensate for
`mitochondrial genome loss in trypanosomes
`Samuel Dean, Matthew K. Gould, Caroline E. Dewar,
`and Achim C. Schnaufer
`
`Treatment with suboptimal doses of raltegravir leads to
`aberrant HIV-1 integrations
`J anani Varaclarajan, Mary Jane McWilliams,
`and Stephen H. Hughes
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`Transgenic mouse model for the study of enterovirus
`71 neuropathogenesis
`Ken Fujii, Noriyo Nagata, Yoko Sato, Kjen
`Chai Ong, Kum Thong Wong, Seiya Yamayoshi,
`Midori Shimanuki, Hiroshi Shitara, Choji Taya,
`and Satoshi Koike
`
`NEUROSCIENCE
`
`Two distinct forms of functional lateralization in the
`human brain
`
`Stephen J. Gotts, Hang Joon Jo, Gregory L. Wallace,
`Ziad S. Saad, Robert W. Cox, and Alex Martin
`
`Neural mechanisms of communicative innovation
`Arjcn Stolk, Lennart Verhagen, Jan-Mathijs Schoffelen,
`Robert Oostenveld, Mark Blokpoel, Peter Hagoort,
`Iris van Rooij, and Ivan Toni
`
`Profoundly different prion diseases in knock-in mice
`carrying single PrP codon substitutions associated with
`human diseases
`Walker 8. Jackson, Andrew W. Borkowski, Nicki E.
`Watson, Oliver D. King, Hemyk Faas, Alan Jasanolf,
`and Susan Lindquist
`
`NRZB subunit of the NMDA glutamate receptor regulates
`appetite in the parabrachial nucleus
`0i Wu, Ruimao Zheng, Dollada Srisai, G. Stanley McKnight,
`and Richard D. Palmiter
`
`Low levels of copper disrupt brain amyloid-p
`homeostasis by altering its production and clearance
`Itender Singh, Abhay P. Sagare, Mireia Coma, David
`Perlmutter, Robert Gelein, Robert D. Bell, Richard J. Deane,
`Elaine Zhong, Margaret Parisi, Joseph Ciszewski, R. Tristan
`Kasper, and Rashid Deane
`
`Disruption of centrifugal inhibition to olfactory bulb
`granule cells impairs olfactory discrimination
`Alexia Nunez—Farm, Robert K, Maurer, Krista Krahe,
`Richard S. Smith, and Ricardo C. Araneda
`
`

`

`14783
`
`14789
`
`14795
`
`E3445
`
`Chronic alcohol produces neuroadaptations to prime
`dorsal striatal learning
`Lauren DePoy, Rachel Daut, Jonathan L Brigman, Kathryn
`MacPherson, Nicole Crowley, Ozge Gunduz-Cinar, Charles L.
`Piekens, Resat Cinar, Lisa M. Saksida, George Kunos,
`David M. Lovinger, Timothy J. Bussey, Marguerite C. Camp,
`and Andrew Holmes
`
`Role of the ectonucleotidase NTPDaseZ in taste
`bud function
`Aurelie Vandenbeuch, Catherine B. Anderson, Jason Parnes,
`Keiichi Enjyoji, Simon C. Robson. Thomas E. Finger,
`and Sue C. Kinnamon
`
`Dorsal periaqueductal gray-amygdala pathway conveys
`both innate and Ieamed ‘lear responses in rats
`Eun Joo Kim, Omer Horovitz, Blake A. Pellrnan, Lancy Mimi
`Tan, Qiuling Li, Gal Richter-Levin, and Jeansok J. Kim
`
`PHARMACOLOGY
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`Stapled a—helical peptide drug development: A potent
`dual inhibitor of MDMZ and MDMX for p53-dependent
`cancer therapy
`Yong S. Chang. Bradford Graves, Vincent Guerlavais,
`Christian Tovar, Kathryn Packman, Kwong-Him To. Karen A.
`Olson, Kamala Kesavan. Pranoti Gangurde, Aditi Mukherjee,
`Theresa Baker, Kmysztof Darlak, Carl Elkin, Zoran Filipovic,
`Farooq Z. Qnreshi, Hongliang Cai, Pamela Berry, Eric
`Feyfant, Xiangguo E. Shi, James Horstick, D. Allen Annis,
`Anthony M. Manning, Nader Fotouhi, Huw Nash,
`Lyubomir T. Vassilev, and Tomi K. Sawyer
`
`PHYSIOLOGY
`
`E3455
`
`Subtype-specific control of P2X receptor channel
`signaling by ATP and Mg2+
`Mufeng Li, Shai D. Silberberg, and Kenton J. Swarlz
`
`14801
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`14807
`
`Hypothalamo-pituitary and immune-dependent adrenal
`regulation during systemic inflammation
`Waldemar Kanczkowski. Vasiieia-Ismini Alexaki, Nguyen
`Tran, Sylvia GroBklaus, Kai Zacharowski, Antoine Martinez,
`Petra Popovics. Norman L. Block, Triantafylios Chavakis,
`Andrew V. Schally. and Stefan R. Bernstein
`
`PLANT BIOLOGY
`
`Salicylic acid 3-hydroxylase regulates Arabidopsis leaf
`longevity by mediating salicylic acid catabolisn'l
`Kewei Zhang, Rayko Halitschke, Changxi Yin, Chang-Jun
`Liu, and Su-Sheng Gan
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`cannecnows -
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`'
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`14813
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`14813
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`BIOPHYSICS AND COMPUTATIONAL BIOLOGY
`
`Structural insights into the proton pumping by unusual
`proteorhodopsin from nonmarine bacteria
`Ivan Gushehin, Pavel Chervakov, Pavel Kuzmichev,
`Alexander N. Popov, Ekaterina Round. Valentin
`Borshchevskiy, Andrii Ishchenko, Lada Petrovskaya,
`Vladimir Chupin, Dmitry A. Dolgikh, Alexander A. Arseniev,
`Mikhail Kirpichnikov, and Valentin Gordeliy
`
`PHYSIOLOGY
`
`Sulfatides are required for renal adaptation to chronic
`metabolic acidosis
`Paula Stettner, Soline Bourgeois, Christian Marsching,
`Milena Traykova-Brauch, Stefan Porubsky, Viola Nordstrc'im.
`Carsten Hopf, Robert Kosters, Roger Sandhoff, Herbert
`Wiegandt, Carsten A Wagner, Hermann-Josef Guinea.
`and Richard Jennemann
`
`is Subscription Form
`
`PNAS
`
`| SeptemherB. 2013 | vo|.1lo | no. 36 l vii
`
`

`

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`JD.
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`this material my bevlotmtzd wows»: law (We 17 u s (odti
`
`
`
`Blockade of Am receptors potently suppresses the
`metastasis of CD731” tumors
`
`Paul A. Beavisa'b, Upulie Divisekeraa’b, Christophe Pageta'b. Melvyn T. Chowa'h, Liza B. Johna'b, Christel Devauda'b,
`Karen Dwyer“. John Stagg“, Mark J. Smytha'b'f'g”, and Phillip K. Darcya'll'L2
`
`a(lancer Immunology Program, Peter MacCaIIum Cancer Centre, East Melbourne, VIC 3002, Australia; bSir Peter MacCaIlum Department of Oncology,
`The University of Melbourne, Parkville, VIC 3010, Australia; ‘St Vincent's Hospital, Immunology Research Centre, Fitzroy, VIC 3065, Australia; dDepartment
`of Medicine, University of Melbourne, Parkville, VIC 3010, Australia; ”Centre de Recherche du Centre Hospitalier de l‘Université de Montreal, Faculté de
`Pharmacie et Institut du Cancer de Montreal, Montreal, QC. Canada HZL 4M1; fImmunology in Cancer and Infection Laboratory, Queensland Institute
`of Medical Research, Herston, QLD 4006, Australia; and 95chool of Medicine, University of Queensland, Herston, QLD 4006, Australia
`
`Edited by Steven A. Rosenberg, ‘National Institutes of Healtthational Cancer Institute, Bethesda. MD. and accepted by the Editorial Board July 23, 2013
`(received for review May S. 2013)
`
`CD73 inhibits antitumor immunity through the activation of aden-
`osine receptors expressed on multiple immune subsets. CD73 also
`enhances tumor metastasis, although the nature of the immune
`subsets and adenosine receptor subtypes involved in this process
`are larger unknown. In this study, we revealed that Aur‘Azg re-
`ceptor antagonists were effective in reducing the metastasis of
`tumors expressing CD73 endogenously (411.2 breast tumors) and
`when CD73 was ectopically expressed (B‘lGF1O melanoma). A35"
`mice were strongly protected against tumor metastasis. indicating
`that host Au receptors enhanced tumor metastasis. A2,. blockade
`enhanced natural killer (NK) cell maturation and cytotoxic function
`in vitro. reduced metastasis in a perform-dependent manner, and
`enhanced NK cell expression of granzyme B in vivo, strongly sug-
`gesting that the antimetastatic effect of A2,. blockade was due to
`enhanced NK cell function. Interestingly, A23 blockade had no
`effect on NK cell cytotoxicity. indicating that an NK cell-indepen-
`dent mechanism also contributed to the increased metastasis of
`CD73" tumors. Our results thus revealed that CD73 promotes tumor
`metastasis through multiple mechanisms, including suppression of
`NK cell function. Furthermore, our data strongly suggest that A” or
`A1,; antagonists may be useful for the treatment of metastatic dis-
`ease. Overall, our study has potential therapeutic implications
`given that ARIA“ receptor antagonist have already entered clin-
`ical trials in other therapeutic settings.
`
`cancer metastasis I immunotherapy I tumor immunosuppression I
`innate immunity
`
`is now well-established that tumors use numerous immuno-
`t
`suppressive mechanisms to facilitate tumor growth (1, 2). Land-
`mark studics by Sitkovsky and colleagues established that one such
`mechanism is the catabolism of extracellular AMP into immuno-
`
`suppressive adenosine {3, 4). Extracellular adenosine is generated
`from adenosine monophosphate (AMP) by the ecloenzyme CD73
`and binds to four known cell surface receptors (A1, Am, A23, and
`A3) that are expressed on multiple immune subsets including T
`cells, natural killer (NK) cells, natural killer T cells, macrophages,
`dendritic cells, and mycloid-derived suppressor cells (MDSCS) (5).
`The A2,, and A23 receptor subtypes are essentially responsible
`for the immunosuppressive effects of adenosine. They share
`a common signaling pathway, both resulting in the activation of
`adenylate cyclase and the accumulation of intracellular CAMP.
`Extracellular AMP can be generated from ATP, an immune-
`activating molecule, by the ectoenzyme CD39. Thus, CD73 and
`CD39 acting in concert can catabolize ATP into adenosine and
`therefore tip the balance from an immunostimulatory to an im-
`munosuppressive microenvironment.
`Physiologically, CD73 is expressed on endothelial cells, epi—
`thelial cells, and subsets of the immune system, including Tregs
`(5). CD73 has been found to be up-regulated in several types of
`cancer and recently it has been shown to be a negative prognostic
`marker in triple negative breast cancer (5, 6). Several preclinical
`studies indicate that targeting the CD73-adenosine pathway may
`
`be beneficial. Proof-of—concept studies demonstrated that target-
`ing CD73 with an anti-CD73—blocking mAb could significantly
`delay tumor growth (7). This was due to enhanced antitumor
`immune T-cell responses as a result of attenuated Aim-mediated
`immunosuppression. Subsequently, both host and tumor-expressed
`CD73 have been shown to be important in the suppression of
`antitumor T-cell responses. Indeed, CD73-deficient mice are re-
`sistant to several transplantable tumors and de novo carcinogen-
`esis (8—11). On the other hand, specific targeting of CD73 on
`tumor cells using shRNA also significantly reduced tumor growth
`and prolonged survival {7, 12).
`Furthermore,
`the knockdown of CD73 expression on tumor
`cells reduced their ability to metastasize (7). However, the mech-
`anism by which CD73 promotes tumor metastasis is unclear.
`Understanding the mechanism by which CD73 increases tumor
`metastasis is vital
`to developing new strategies that target
`metastasis because there are no anti-CD73 mAbs currently in
`clinical development. The clinical relevance of this pathway is
`underlined by the correlation between CD73 expression and
`an increased risk of metastasis in breast cancer {13, 14). Thus,
`the CD73 pathway has been proposed as a novel target for
`immunotherapy (15).
`In this current study, we found that CD73 promoted metas-
`tasis through the activation of both A”, and A23 receptors. A2,».
`blockade resulted in enhanced NK—oell activity in vitro and in
`vivo and reduced metastasis in a perform—dependent manner.
`Titus, Am or A2]; antagonists, which have already been used in
`clinical trials in other disease settings (16), are promising for the
`treatment of cancer metastasis for which there is a lack of ef-
`
`fective options available.
`Results
`
`Tumor-CD73 Expression Promotes in Vivo Metastasis. Previous inves-
`tigations in our laboratory suggested that the expression of CD73
`on tumor cells could positively regulate metastasis (7}. To in-
`vestigate whether CD73 expression was sufficient to increase the
`migratory properties of tumor cells, AT-3 breast adenocarcinoma
`and B16F10 melanoma cells (both CD73') were transduoed with
`retrovirus-encoding murine CD73 cDNA, CD73 expression on
`transduced AT—3 and BlfiFlO cells was at a level equivalent to
`4T1.2 breast carcinoma cells, a metastatic cell line expressing high
`levels of CD73 (Fig 51A) {7). Eclopically expressed CD73 was
`
`Author contributions: P.A.B., J.S., MJ.S., and P.l(.D. designed research; P.A.B., U.D., C.P.,
`M.T.C.. L.B.J., C.D., and P.K.D. performed research: K.D. and MJS. contribu