Tristetraprolin inhibits HIV-1 production by binding to genomic RNA.
HIV-1 genome has an AU-rich sequence and requires rapid nuclear export by Rev activity to prevent multiple splicing. HIV-1 infection occurs in activated CD4(+) T cells where the decay of mRNAs of cytokines and chemokines is regulated by the binding of AU-rich elements to the mRNA-destabilizing protein tristetraprolin. We here investigated the influence of tristetraprolin on the replication of HIV-1. Treatment of siRNA against tristetraprolin in a latently HIV-1 infected cell line increases HIV-1 production following stimulation. A chloramphenicol acetyltransferase and luciferase assay revealed that exogenous tristetraprolin reduced HIV-1 virion production and in contrast increased the multiply spliced products. Furthermore, quantitative RT-PCR analysis showed tristetraprolin increases the ratio of multiple-spliced RNAs to un-, single-spliced RNA. Moreover, an electrophoretic mobility shift assay showed that tristetraprolin binds to synthesized HIV-1 RNA with AU-rich sequence but not to RNA with less AU sequence. These results suggest that tristetraprolin is a regulator of HIV-1 replication and enhances splicing by direct binding to AU-rich sequence of HIV-1 RNAs.
Maeda
Masae
M
Department of Pathology, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku-ku, Tokyo 162-8640, Japan.
Sawa
Hirofumi
H
Tobiume
Minoru
M
Tokunaga
Kenzo
K
Hasegawa
Hideki
H
Ichinohe
Takeshi
T
Sata
Tetsutaro
T
Moriyama
Masami
M
Hall
William W
WW
Kurata
Takeshi
T
Takahashi
Hidehiro
H
D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000032:analysis / Q000235:genetics; D024202:Electrophoretic Mobility Shift Assay; D020868:Gene Silencing; D016679:Genome, Viral; D015497:HIV-1 / Q000235:genetics / Q000502:physiology*; D006801:Humans; D008156:Luciferases / Q000032:analysis / Q000235:genetics; D011485:Protein Binding; D012326:RNA Splicing; D034741:RNA, Small Interfering / Q000378:metabolism; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D051816:Tristetraprolin / Q000037:antagonists & inhibitors / Q000378:metabolism*; D014779:Virus Replication
D034741:RNA, Small Interfering; D012367:RNA, Viral; D051816:Tristetraprolin; D008156:Luciferases; D015500:Chloramphenicol O-Acetyltransferase