Identification and characterization of PWWP domain residues critical for LEDGF/p75 chromatin binding and human immunodeficiency virus type 1 infectivity.
Lens epithelium-derived growth factor (LEDGF)/p75 functions as a bimodal tether during lentiviral DNA integration: its C-terminal integrase-binding domain interacts with the viral preintegration complex, whereas the N-terminal PWWP domain can bind to cellular chromatin. The molecular basis for the integrase-LEDGF/p75 interaction is understood, while the mechanism of chromatin binding is unknown. The PWWP domain is homologous to other protein interaction modules that together comprise the Tudor clan. Based on primary amino acid sequence and three-dimensional structural similarities, 24 residues of the LEDGF/p75 PWWP domain were mutagenized to garner essential details of its function during human immunodeficiency virus type 1 (HIV-1) infection. Mutating either Trp-21 or Ala-51, which line the inner wall of a hydrophobic cavity that is common to Tudor clan members, disrupts chromatin binding and virus infectivity. Consistent with a role for chromatin-associated LEDGF/p75 in stimulating integrase activity during infection, recombinant W21A protein is preferentially defective for enhancing integration into chromatinized target DNA in vitro. The A51P mutation corresponds to the S270P change in DNA methyltransferase 3B that causes human immunodeficiency, centromeric instability, and facial anomaly syndrome, revealing a critical role for this amino acid position in the chromatin binding functions of varied PWWP domains. Our results furthermore highlight the requirement for a conserved Glu in the hydrophobic core that mediates interactions between other Tudor clan members and their substrates. This initial systematic mutagenesis of a PWWP domain identifies amino acid residues critical for chromatin binding function and the consequences of their changes on HIV-1 integration and infection.
Shun
Ming-Chieh
MC
Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute and Division of AIDS, Harvard Medical School, Boston, MA 02115, USA.
Botbol
Yaïr
Y
Li
Xiang
X
Di Nunzio
Francesca
F
Daigle
Janet E
JE
Yan
Nan
N
Lieberman
Judy
J
Lavigne
Marc
M
Engelman
Alan
A
D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry / Q000502:physiology*; D000818:Animals; D002460:Cell Line; D002843:Chromatin / Q000378:metabolism*; D015497:HIV-1 / Q000472:pathogenicity*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000737:chemistry / Q000502:physiology*; D051379:Mice; D017434:Protein Structure, Tertiary; D014157:Transcription Factors / Q000737:chemistry / Q000502:physiology*; D016662:Virus Integration
D048868:Adaptor Proteins, Signal Transducing; D002843:Chromatin; D036341:Intercellular Signaling Peptides and Proteins; C504649:PSIP1 protein, human; D014157:Transcription Factors; C119380:lens epithelium-derived growth factor
Proc Natl Acad Sci U S A. 2006 Jan 31;103(5):1498-503
16428288
J Virol. 2007 Jan;81(1):166-72
17035312
Nat Struct Mol Biol. 2005 Jun;12(6):526-32
15895093
PLoS Pathog. 2008 Mar 21;4(3):e1000027
18369476
Nucleic Acids Res. 2006 Jan 1;34(Database issue):D247-51
16381856
J Allergy Clin Immunol. 2000 Jun;105(6 Pt 1):1211-20
10856157
J Gen Virol. 2006 May;87(Pt 5):1339-1347
16603537
J Virol. 2004 Sep;78(17):9524-37
15308744
J Virol. 2006 Feb;80(4):1886-96
16439544
J Biol Chem. 2003 Aug 29;278(35):33528-39
12796494
Virology. 2007 Jan 5;357(1):79-90
16959283
Cancer Cell. 2008 Jul 8;14(1):36-46
18598942
Front Biosci. 2004 Sep 01;9:3187-208
15353349
Nat Med. 2005 Dec;11(12):1287-9
16311605
Cell. 2002 Aug 23;110(4):521-9
12202041
BMC Mol Biol. 2007 Oct 31;8:101
17974029
Am J Med Genet. 2002 Sep 15;112(1):31-7
12239717
J Mol Biol. 2003 Jul 11;330(3):571-6
12842472
PLoS Pathog. 2008 Mar 28;4(3):e1000046
18369482
Antimicrob Agents Chemother. 2002 Jun;46(6):1896-905
12019106
J Mol Biol. 2006 Jan 20;355(3):379-94
16318853
Virology. 2006 Mar 15;346(2):415-26
16337983
J Virol. 2007 Apr;81(8):3969-79
17267486
Proc Natl Acad Sci U S A. 2005 Nov 29;102(48):17308-13
16260736
J Virol. 2008 Feb;82(3):1360-7
18032509
J Cell Sci. 2005 Apr 15;118(Pt 8):1733-43
15797927
J Virol. 2006 Dec;80(23):11498-509
16987986
J Biol Chem. 2004 Aug 6;279(32):33421-9
15163664
PLoS Pathog. 2006 Jun;2(6):e60
16789841
Nucleic Acids Res. 2006 Mar 20;34(5):1653-65
16549878
PLoS One. 2007 Dec 19;2(12):e1340
18092005
Development. 2008 Jun;135(11):1935-46
18469222
J Biol Chem. 2003 Jan 3;278(1):372-81
12407101
Cell Mol Life Sci. 2008 May;65(9):1403-24
18264802
J Virol. 2004 Nov;78(21):11656-63
15479807
FEBS Lett. 2000 May 4;473(1):1-5
10802047
Science. 2006 Oct 20;314(5798):461-4
16959972
Trends Biochem Sci. 2003 Feb;28(2):69-74
12575993
J Biol Chem. 2004 Nov 19;279(47):48883-92
15371438
J Biol Chem. 2005 Jul 8;280(27):25517-23
15855167
FEBS J. 2008 Apr;275(7):1411-1426
18331345
PLoS Biol. 2004 Dec;2(12):e423
15550989
J Virol. 2005 Apr;79(8):5211-4
15795304
J Virol. 2007 Jun;81(12):6731-41
17409138
Nat Struct Biol. 2002 Mar;9(3):217-24
11836534
J Mol Biol. 2007 Feb 2;365(5):1480-92
17137594
Cell Tissue Res. 2001 Jul;305(1):107-14
11512661
J Mol Biol. 2006 Jul 21;360(4):760-73
16793062
Science. 2002 Mar 15;295(5562):2080-3
11859155
Virology. 2007 Mar 30;360(1):1-5
17258258
Nucleic Acids Res. 2007;35(1):113-24
17158150
J Virol. 1995 May;69(5):2729-36
7535863
Genes Dev. 2007 Jul 15;21(14):1767-78
17639082
Protein Sci. 2006 Feb;15(2):314-23
16384999
PLoS Pathog. 2007 Mar;3(3):e47
17397262
Nucleic Acids Res. 2008 Mar;36(4):1237-46
18174227
Biochem Biophys Res Commun. 1997 Sep 8;238(1):26-32
9299445
J Biol Chem. 1994 Oct 7;269(40):25143-9
7929202
Science. 2003 Jun 13;300(5626):1749-51
12805549
J Virol. 2004 Dec;78(23):12735-46
15542626
Protein Sci. 2005 Mar;14(3):756-64
15689505
J Virol. 2002 Nov;76(21):10598-607
12368302
Nat Rev Microbiol. 2005 Nov;3(11):848-58
16175173
Nature. 2002 Mar 7;416(6876):103-7
11882902
J Biol Chem. 2004 Jun 11;279(24):25447-54
14998998
PLoS Biol. 2004 Aug;2(8):E234
15314653
Nat Struct Biol. 2001 Jan;8(1):27-31
11135666
Proc Natl Acad Sci U S A. 2005 Apr 26;102(17):6103-7
15802467
Biochemistry. 2007 Mar 13;46(10):2899-908
17298035