AKAP149 binds to HIV-1 reverse transcriptase and is involved in the reverse transcription.
Like all retroviruses, human immunodeficiency virus type 1 (HIV-1) undergoes reverse transcription during its replication cycle. The cellular cofactors potentially involved in this process still remain to be identified. We show here that A-kinase anchoring protein 149 (AKAP149) interacts with HIV-1 reverse transcriptase (RT) in both the yeast two-hybrid system and human cells. The AKAP149 binding site has been mapped to the RNase H domain of HIV-1 RT. AKAP149 silencing by RNA interference in HIV-1-infected cells inhibited viral replication at the reverse transcription step. We selected single-point mutants of RT defective for AKAP149 binding and demonstrated that mutant G462R, despite retaining significant intrinsic RT activity in vitro, failed to carry out HIV-1 reverse transcription correctly in infected cells. This suggests that the interaction between RT and AKAP149 in infected cells may play an important role in HIV-1 reverse transcription.
Lemay
Julie
J
Institut Cochin, Université Paris Descartes, CNRS (UMR 8104), Paris, France.
Maidou-Peindara
Priscilla
P
Cancio
Reynel
R
Ennifar
Eric
E
Coadou
Gaël
G
Maga
Giovanni
G
Rain
Jean-Christophe
JC
Benarous
Richard
R
Liu
Lang Xia
LX
D054758:A Kinase Anchor Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D054303:HIV Reverse Transcriptase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015497:HIV-1 / Q000201:enzymology / Q000235:genetics; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016296:Mutagenesis; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D034622:RNA Interference; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D048348:Reverse Transcription / Q000502:physiology*; D020798:Two-Hybrid System Techniques; D014771:Virion / Q000378:metabolism; D014779:Virus Replication / Q000502:physiology
D054758:A Kinase Anchor Proteins; C105137:AKAP1 protein, human; D011993:Recombinant Fusion Proteins; C514824:reverse transcriptase, Human immunodeficiency virus 1; D054303:HIV Reverse Transcriptase