title: A carboxy-terminally truncated human CPSF6 lacking residues encoded by exon 6 inhibits HIV-1 cDNA synthesis and promotes capsid disassembly.
abstract: Since HIV-1 replication is modulated at multiple stages by host cell factors, identification and characterization of those host cell factors are expected to contribute to the development of novel anti-HIV therapeutics. Previous studies showed that a C-terminally truncated cytosolic form of cleavage and polyadenylation-specific factor 6 (CPSF6-358) inhibits HIV-1 infection through interference with HIV-1 trafficking to the nucleus. Here we identified and characterized a different configuration of C-terminally truncated human CPSF6 (hCPSF6-375) through cDNA expression cloning coupled with ganciclovir-mediated lethal selection. Notably, hCPSF6-375, but not mouse CPSF6-358 (mCPSF6-358) as previously reported, remarkably interfered with viral cDNA synthesis after HIV-1 infection. Moreover, we found that hCPSF6-375 aberrantly accelerated the disassembly of the viral capsid in target cells, while CPSF6-358 did not. Sequence comparison of CPSF6-375 and CPSF6-358 cDNAs showed a lack of exon 6 and additional coding sequence for 54 amino acid residues in the C terminus of hCPSF6-375. Mutational analyses revealed that the residues encoded by exon 6, but not the C-terminal 54 residues in hCPSF6-375, is responsible for impaired viral cDNA synthesis by hCPSF6-375. This is the first report demonstrating a novel mode of HIV-1 inhibition by truncated forms of CPSF6 that involves rapid capsid disassembly and inhibition of viral cDNA synthesis. These findings could facilitate an increased understanding of viral cDNA synthesis in light of the viral capsid disassembly.
authors: Department of Molecular Virology, Tokyo Medical and Dental University, Tokyo, Japan.
authors: Takanori
authors: T
authors: Hori
authors: Hiroaki
authors: H
authors: Takeuchi
authors: Hideki
authors: H
authors: Saito
authors: Ryuta
authors: R
authors: Sakuma
authors: Yoshio
authors: Y
authors: Inagaki
authors: Shoji
authors: S
authors: Yamaoka
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002213:Capsid / Q000378:metabolism*; D039223:Cleavage And Polyadenylation Specificity Factor / Q000235:genetics; D004252:DNA Mutational Analysis; D018076:DNA, Complementary / Q000096:biosynthesis*; D005822:Genetic Vectors / Q000235:genetics; D057809:HEK293 Cells; D015497:HIV-1 / Q000235:genetics* / Q000502:physiology; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D014779:Virus Replication / Q000235:genetics* / Q000502:physiology; D039221:mRNA Cleavage and Polyadenylation Factors / Q000235:genetics* / Q000378:metabolism*
chemical_list: C529056:CPSF6 protein, mouse; D039223:Cleavage And Polyadenylation Specificity Factor; D018076:DNA, Complementary; C450562:cleavage factor Im, human; D039221:mRNA Cleavage and Polyadenylation Factors
references: FEBS Lett. 2002 Nov 20;531(3):553-60
references: 12435610
references: FEBS Lett. 2008 Dec 10;582(29):4053-8
references: 18996376
references: Cancer Res. 1986 Oct;46(10):5276-81
references: 3019523
references: J Virol. 2002 Jun;76(11):5667-77
references: 11991995
references: Cell Host Microbe. 2010 Mar 18;7(3):221-33
references: 20227665
references: J Virol. 2001 Oct;75(19):9357-66
references: 11533199
references: Trends Microbiol. 2004 Apr;12(4):170-7
references: 15051067
references: Science. 2008 Feb 15;319(5865):921-6
references: 18187620
references: Proc Natl Acad Sci U S A. 2009 Mar 17;106(11):4071-8
references: 19237569
references: J Gen Virol. 2007 Jan;88(Pt 1):207-216
references: 17170453
references: J Biol Chem. 2001 Jul 20;276(29):27519-26
references: 11375395
references: J Biol Chem. 2005 Jan 7;280(1):375-82
references: 15528183
references: J Biol Chem. 2004 Aug 20;279(34):35788-97
references: 15169763
references: Cell. 2008 Oct 3;135(1):49-60
references: 18854154
references: J Virol. 2003 May;77(9):5439-50
references: 12692245
references: Jpn J Infect Dis. 2008 Jul;61(4):279-85
references: 18653969
references: J Virol. 2003 Apr;77(7):4409-14
references: 12634398
references: PLoS Pathog. 2012;8(8):e1002896
references: 22956906
references: Retrovirology. 2012 Jan 06;9:3
references: 22225545
references: J Virol. 2007 Aug;81(15):8080-90
references: 17522232
references: Cell Host Microbe. 2008 Nov 13;4(5):495-504
references: 18976975
references: Mol Cell. 2006 Aug;23(4):597-605
references: 16916646
references: Nat Rev Microbiol. 2007 Apr;5(4):253-63
references: 17325726
references: Proc Natl Acad Sci U S A. 2006 Apr 4;103(14):5514-9
references: 16540544
references: J Virol. 1995 Sep;69(9):5883-9
references: 7637036
references: J Virol. 2004 Oct;78(20):11352-9
references: 15452256
references: J Virol. 2012 Apr;86(7):3851-60
references: 22301135
references: FEBS Lett. 2002 Dec 4;532(1-2):45-51
references: 12459460
references: J Biol Chem. 2003 Sep 19;278(38):36005-12
references: 12840022
references: Gene Ther. 2000 Jun;7(12):1063-6
references: 10871756
references: Blood. 2008 May 15;111(10):5118-29
references: 18305221
references: Proc Natl Acad Sci U S A. 1996 Jul 23;93(15):7955-60
references: 8755584
references: J Virol. 2005 May;79(9):5870-4
references: 15827203
references: J Virol. 1988 Jan;62(1):139-47
references: 3257102
references: Nature. 2004 Feb 26;427(6977):848-53
references: 14985764
references: J Virol. 2000 Dec;74(23):11055-66
references: 11070000
references: J Virol. 2001 Aug;75(16):7252-65
references: 11461998
references: J Virol. 2010 May;84(10):5181-90
references: 20219923