title: HIV-1 Gag associates with specific uropod-directed microdomains in a manner dependent on its MA highly basic region.
abstract: In polarized T cells, HIV-1 Gag localizes to a rear-end protrusion known as the uropod in a multimerization-dependent manner. Gag-laden uropods participate in formation of virological synapses, intercellular contact structures that play a key role in cell-to-cell HIV-1 transmission. Our previous observations suggest that Gag associates with uropod-directed microdomains (UDMs) that eventually comigrate with Gag to the uropod over the cell surface. However, the nature of Gag multimerization required for this movement, the composition of the UDMs, and the molecular determinants for Gag association with these microdomains remain unknown. In this study, we found that Gag multimerization prior to budding but beyond dimerization is necessary for Gag localization to the uropods, indicating that uropod localization occurs early in the assembly process. We also found that prior to membrane curvature, Gag multimers associate with a specific subset of UDMs containing PSGL-1, CD43, and CD44 but not ICAM-1, ICAM-3, or CD59. Notably, upon association, Gag excludes ICAM-3 from this subset of UDMs, revealing an active and selective reorganization of these microdomains by Gag. This specific association between Gag and UDMs is dependent on the highly basic region (HBR) in the Gag matrix (MA) domain. The overall positive charge of the HBR was needed for the interaction with the specific UDM subset, while the exact HBR sequence was not, unlike that seen for MA binding to the plasma membrane phospholipid phosphatidylinositol-(4,5)-bisphosphate [PI(4,5)P2]. Taken together, these findings revealed that HIV-1 Gag associates with specific microdomains present in polarized T cells in an MA-dependent manner, which results in modification of the microdomain constituents.
authors.affiliation: Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, Michigan, USA.
authors.forename: G Nicholas
authors.initials: GN
authors.lastname: Llewellyn
authors.forename: Jonathan R
authors.initials: JR
authors.lastname: Grover
authors.forename: Balaji
authors.initials: B
authors.lastname: Olety
authors.forename: Akira
authors.initials: A
authors.lastname: Ono
mesh_terms: D015815:Cell Adhesion Molecules / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D016764:Cell Polarity; D015658:HIV Infections / Q000378:metabolism* / Q000503:physiopathology / Q000821:virology; D015497:HIV-1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D021962:Membrane Microdomains / Q000378:metabolism / Q000821:virology*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D013601:T-Lymphocytes / Q000166:cytology* / Q000378:metabolism / Q000821:virology*; D054301:gag Gene Products, Human Immunodeficiency Virus / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
chemical_list: D015815:Cell Adhesion Molecules; D054301:gag Gene Products, Human Immunodeficiency Virus
references.citation: J Virol. 1998 Mar;72(3):1782-9
references.pmid: 9499028
references.citation: J Virol. 2001 Sep;75(17):7913-24
references.pmid: 11483736
references.citation: Nat Commun. 2012;3:1320
references.pmid: 23271654
references.citation: Virology. 2007 Aug 1;364(2):383-94
references.pmid: 17434553
references.citation: J Virol. 2005 Nov;79(22):14131-40
references.pmid: 16254348
references.citation: Proc Natl Acad Sci U S A. 2003 Mar 4;100(5):2604-9
references.pmid: 12601158
references.citation: J Virol. 2011 Oct;85(20):10851-60
references.pmid: 21813603
references.citation: Nat Rev Mol Cell Biol. 2007 Mar;8(3):185-94
references.pmid: 17318224
references.citation: J Exp Med. 2006 Nov 27;203(12):2749-61
references.pmid: 17116735
references.citation: J Cell Biol. 2008 Nov 3;183(3):527-42
references.pmid: 18955551
references.citation: J Virol. 2012 Mar;86(5):2447-58
references.pmid: 22171268
references.citation: Proc Natl Acad Sci U S A. 2006 Jul 25;103(30):11364-9
references.pmid: 16840558
references.citation: J Cell Biol. 2006 Mar 13;172(6):923-35
references.pmid: 16533950
references.citation: Nat Rev Mol Cell Biol. 2009 May;10(5):353-9
references.pmid: 19373240
references.citation: J Mol Biol. 2011 Jul 22;410(4):525-33
references.pmid: 21762798
references.citation: Science. 2003 Mar 14;299(5613):1713-6
references.pmid: 12589003
references.citation: Biochem J. 2009 May 13;420(2):133-54
references.pmid: 19426143
references.citation: J Virol. 1999 Oct;73(10):8527-40
references.pmid: 10482606
references.citation: J Exp Med. 2004 Jan 19;199(2):283-93
references.pmid: 14734528
references.citation: Science. 1997 Oct 31;278(5339):849-53
references.pmid: 9346481
references.citation: Nat Immunol. 2004 Dec;5(12):1235-42
references.pmid: 15516925
references.citation: J Virol. 1993 Nov;67(11):6387-94
references.pmid: 8411340
references.citation: Biochemistry. 2006 Apr 4;45(13):4077-83
references.pmid: 16566581
references.citation: Proc Natl Acad Sci U S A. 2010 Nov 16;107(46):19850-5
references.pmid: 21045134
references.citation: J Mol Biol. 2011 Jul 22;410(4):512-24
references.pmid: 21762797
references.citation: J Virol. 1998 May;72(5):4403-7
references.pmid: 9557731
references.citation: Nature. 2011 Oct 23;479(7374):552-5
references.pmid: 22020284
references.citation: Proc Natl Acad Sci U S A. 2004 Jan 13;101(2):517-22
references.pmid: 14699046
references.citation: EMBO J. 1999 Feb 1;18(3):501-11
references.pmid: 9927410
references.citation: Nat Rev Microbiol. 2005 Aug;3(8):643-55
references.pmid: 16064056
references.citation: J Virol. 2009 Dec;83(23):12196-203
references.pmid: 19776118
references.citation: J Virol. 1999 May;73(5):4136-44
references.pmid: 10196310
references.citation: J Cell Biol. 2008 Nov 3;183(3):375-6
references.pmid: 18981226
references.citation: Eur J Immunol. 2002 Jun;32(6):1560-6
references.pmid: 12115638
references.citation: Immunity. 2006 Dec;25(6):989-1001
references.pmid: 17112751
references.citation: Proc Natl Acad Sci U S A. 1990 Jan;87(2):523-7
references.pmid: 2405382
references.citation: J Virol. 1995 Oct;69(10):6487-97
references.pmid: 7666550
references.citation: J Virol. 2009 Jul;83(14):7322-36
references.pmid: 19403686
references.citation: J Virol. 2000 Jun;74(12):5395-402
references.pmid: 10823843
references.citation: J Virol. 2008 Nov;82(22):11228-38
references.pmid: 18799574
references.citation: J Cell Biol. 1995 Oct;131(2):495-508
references.pmid: 7593174
references.citation: Science. 2002 May 3;296(5569):913-6
references.pmid: 11988576
references.citation: Science. 2009 Mar 27;323(5922):1743-7
references.pmid: 19325119
references.citation: J Virol. 1986 Aug;59(2):284-91
references.pmid: 3016298
references.citation: J Cell Biol. 2006 Jun 5;173(5):795-807
references.pmid: 16735575
references.citation: Blood. 2000 Apr 1;95(7):2413-9
references.pmid: 10733515
references.citation: J Virol. 2011 Jul;85(14):7419-35
references.pmid: 21543480
references.citation: J Leukoc Biol. 1993 Jan;53(1):1-10
references.pmid: 7678845
references.citation: Physiology (Bethesda). 2011 Aug;26(4):236-51
references.pmid: 21841072
references.citation: J Virol. 1999 Mar;73(3):2270-9
references.pmid: 9971810
references.citation: J Virol. 2005 Feb;79(3):1803-12
references.pmid: 15650204
references.citation: J Biol Chem. 2001 Mar 16;276(11):7974-84
references.pmid: 11113129
references.citation: Science. 2002 Jun 7;296(5574):1869-73
references.pmid: 12016203
references.citation: Mol Biol Int. 2012;2012:979765
references.pmid: 22830021
references.citation: Retrovirology. 2009 Jul 14;6:64
references.pmid: 19602278
references.citation: J Cell Biol. 2005 Jul 18;170(2):317-25
references.pmid: 16027225
references.citation: J Virol. 2011 Oct;85(19):9749-66
references.pmid: 21813604
references.citation: J Gen Virol. 1996 Apr;77 ( Pt 4):743-51
references.pmid: 8627263
references.citation: Proc Natl Acad Sci U S A. 2002 Oct 1;99(20):13037-42
references.pmid: 12244217
references.citation: Proc Natl Acad Sci U S A. 2001 Nov 20;98(24):13925-30
references.pmid: 11717449
references.citation: PLoS Biol. 2007 Jun;5(6):e158
references.pmid: 17550307
references.citation: Biochemistry. 2006 Dec 26;45(51):15224-31
references.pmid: 17176044
references.citation: J Mol Biol. 2007 Feb 16;366(2):574-85
references.pmid: 17188710
references.citation: Virology. 1998 Nov 10;251(1):141-57
references.pmid: 9813210
references.citation: Retrovirology. 2009 Mar 11;6:28
references.pmid: 19284574
references.citation: J Virol. 2013 Apr;87(8):4650-64
references.pmid: 23408603
references.citation: Proc Natl Acad Sci U S A. 2004 Oct 12;101(41):14889-94
references.pmid: 15465916
references.citation: J Virol. 2008 Mar;82(5):2405-17
references.pmid: 18094158
references.citation: Curr Top Microbiol Immunol. 2009;339:85-102
references.pmid: 20012525
references.citation: Cell Mol Life Sci. 2005 Oct;62(19-20):2228-40
references.pmid: 16091845
references.citation: Nature. 1965 Feb 27;205:887-8
references.pmid: 14293154
references.citation: J Cell Biol. 2008 Aug 25;182(4):765-76
references.pmid: 18710926
references.citation: J Virol. 2007 Jun;81(12):6434-45
references.pmid: 17392361
references.citation: Trends Biochem Sci. 2011 Nov;36(11):604-15
references.pmid: 21917465
references.citation: J Virol. 2000 Mar;74(6):2855-66
references.pmid: 10684302
references.citation: J Virol. 2005 Sep;79(18):12088-94
references.pmid: 16140785
references.citation: J Virol. 2000 Apr;74(7):3046-57
references.pmid: 10708419
references.citation: J Virol. 1996 Dec;70(12):8540-8
references.pmid: 8970978
references.citation: J Virol. 1993 Apr;67(4):2182-90
references.pmid: 8445728
references.citation: Biol Cell. 2010 Mar 25;102(6):335-50
references.pmid: 20356318
references.citation: J Virol. 2010 Apr;84(7):3516-27
references.pmid: 20089656
references.citation: J Virol. 2007 Nov;81(22):12582-95
references.pmid: 17728240
references.citation: Nature. 2005 Dec 1;438(7068):605-11
references.pmid: 16319880
references.citation: J Virol. 2000 Apr;74(7):3264-72
references.pmid: 10708443
references.citation: Science. 2010 Jan 1;327(5961):46-50
references.pmid: 20044567
references.citation: Proc Natl Acad Sci U S A. 1996 Apr 2;93(7):3099-104
references.pmid: 8610175
references.citation: Virology. 2009 May 10;387(2):341-52
references.pmid: 19285328
references.citation: Proc Natl Acad Sci U S A. 2010 Jan 26;107(4):1600-5
references.pmid: 20080620
references.citation: J Virol. 2003 May;77(9):5439-50
references.pmid: 12692245
references.citation: J Virol. 2007 Aug;81(15):7873-84
references.pmid: 17522207
references.citation: PLoS Pathog. 2010 Feb 26;6(2):e1000788
references.pmid: 20195464
references.citation: PLoS Pathog. 2010 Oct 28;6(10):e1001167
references.pmid: 21060818
references.citation: Traffic. 2010 Nov;11(11):1401-14
references.pmid: 20727121
references.citation: Proc Natl Acad Sci U S A. 1989 Aug;86(15):5781-5
references.pmid: 2788277
references.citation: J Biol Chem. 2011 Dec 23;286(51):44162-44176
references.pmid: 21865156
references.citation: J Virol. 2010 Apr;84(7):3248-58
references.pmid: 20089647
references.citation: Nature. 2012 Oct 11;490(7419):283-7
references.pmid: 22854780
references.citation: J Virol. 1994 Apr;68(4):2556-69
references.pmid: 8139035
references.citation: J Mol Biol. 2007 Jan 19;365(3):799-811
references.pmid: 17098251
references.citation: J Virol. 2009 Mar;83(5):2216-25
references.pmid: 19073719
references.citation: Proc Natl Acad Sci U S A. 2009 Aug 11;106(32):13301-6
references.pmid: 19620730
references.citation: Cell Host Microbe. 2009 Jun 18;5(6):550-8
references.pmid: 19527882
references.citation: Proc Natl Acad Sci U S A. 2001 Apr 24;98(9):5246-51
references.pmid: 11320254
references.citation: J Virol. 2006 Mar;80(6):2884-93
references.pmid: 16501097
references.citation: J Cell Biol. 1997 Apr 21;137(2):493-508
references.pmid: 9128258
references.citation: J Virol. 2009 Jun;83(12):6234-46
references.pmid: 19369333