• ホーム
  • 検索結果
  • Binding of the eukaryotic translation elongation factor 1A with the 5'UTR of HIV-1 genomic RNA is important for reverse transcription.

Binding of the eukaryotic translation elongation factor 1A with the 5'UTR of HIV-1 genomic RNA is important for reverse transcription.

Virology journal (2015-08-06)
Dongsheng Li, Ting Wei, Hongping Jin, Amanda Rose, Rui Wang, Min-Hsuan Lin, Kirsten Spann, David Harrich
要旨

The cellular protein eukaryotic translation elongation factor 1A (eEF1A) binds to aminoacylated transfer RNAs and delivers them to the ribosome during translation. eEF1A also binds to RNA secondary structures present in genomes of several RNA viruses and plays important roles in their replication. As a RNA binding protein, whether eEF1A can bind with HIV-1 genomic RNA has not been investigated and was the aim of the study. RNA-protein interaction was determined by reversible crosslink co-immunoprecipitation (RC-Co-IP) and biolayer Interferometry assay (BLI). eEF1A binding region within RNA was mapped by deletion and mutation analysis. Virus with genomic RNA mutations were examined for eEF1A-RT interaction by proximity ligation assay, for reverse transcription by qPCR and for replication by CAp24 ELISA in cells. The interaction of eEF1A with 5'UTR of HIV-1 genomic RNA was detected in cells and in vitro. Truncation and substitution mutations in the 5'UTR RNA demonstrated that a stem-loop formed by nucleotides 142 to 170, which encompass a reported tRNA anticodon-like-element, binds to eEF1A. Mutations that altered the stem-loop structure by changing two highly conserved sequence clusters in the stem-loop region result in reduction of the interaction with eEF1A in vitro. HIV-1 virus harbouring the same 5'UTR mutations significantly reduced the interaction of eEF1A with HIV-1 reverse transcription complex (RTC), reverse transcription and replication. eEF1A interacts with 5'UTR of HIV-1 genomic RNA and the interaction is important for late DNA synthesis in reverse transcription.

材料
製品番号
ブランド
製品内容

Sigma-Aldrich
Monoclonal Anti-EEF1A1 antibody produced in mouse, clone EF48, purified from hybridoma cell culture
Sigma-Aldrich
Anti-EEF1A1 antibody produced in rabbit, IgG fraction of antiserum

ソーシャルメディア

LinkedIn icon
Twitter icon
Facebook Icon
Instagram Icon

Merck

研究、開発、製造

私たちは、バイオテクノロジーと医薬品の研究&開発および製造のソリューションとサービスを提供する、グローバルなライフサイエンス業界のリーディングサプライヤーです。

© 2021 Merck KGaA, Darmstadt, Germany and/or its affiliates.All Rights Reserved.

当ウェブサイトのいかなる内容についても、許可なく複写することを禁じます。