Specific interaction of HeLa cell proteins with coxsackievirus B3 3′UTR

La autoantigen binds the 3′ and 5′UTR independently of the poly(A) tail

Paul Cheung, Travis Lim, Ji Yuan, Mary Zhang, David Chau, Bruce Mcmanus, Decheng Yang

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Coxsackievirus B3 (CVB3) is a positive, single-stranded RNA virus. The secondary structure of the 3′ untranslated region (3′UTR) of CVB3 RNA consists of three stem-loops and is followed by a poly(A) tail sequence. These stem-loop structures have been suggested to participate in the regulation of viral replication through interaction with cellular proteins that are yet to be identified. In this study, by competitive UV cross-linking using mutated 3′UTR probes we have demonstrated that the poly(A) tail is essential for promoting HeLa cell protein interactions with the 3′UTR because deletion of this sequence abolished most of the protein interactions. Unexpectedly, mutations that disrupted the tertiary loop-loop interactions without affecting the stem-loops did not apparently affect these protein interactions, indicating that secondary structure rather than the high-order structure may play a major role in recruiting these RNA binding proteins. Among the observed 3′UTR RNA binding proteins, we have confirmed a 52 kDa protein as the human La autoantigen by using purified recombinant protein and a polyclonal La antibody. This protein can interact with both the 3′ and 5′UTRs independently of the poly(A) tail. Further analysis by two-stage UV cross-linking, we found that the 3′ and 5′UTR sequences may share the same binding site on the La protein.

Original languageEnglish (US)
Pages (from-to)1705-1715
Number of pages11
JournalCellular Microbiology
Volume9
Issue number7
DOIs
StatePublished - Jul 1 2007

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Enterovirus
Autoantigens
3' Untranslated Regions
HeLa Cells
Messenger RNA
Proteins
RNA-Binding Proteins
Untranslated RNA
RNA
Sequence Deletion
RNA Viruses
Recombinant Proteins
Cell Communication
Viruses
Binding Sites
Mutation
Antibodies

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Virology

Cite this

Specific interaction of HeLa cell proteins with coxsackievirus B3 3′UTR : La autoantigen binds the 3′ and 5′UTR independently of the poly(A) tail. / Cheung, Paul; Lim, Travis; Yuan, Ji; Zhang, Mary; Chau, David; Mcmanus, Bruce; Yang, Decheng.

In: Cellular Microbiology, Vol. 9, No. 7, 01.07.2007, p. 1705-1715.

Research output: Contribution to journalArticle

Cheung, Paul ; Lim, Travis ; Yuan, Ji ; Zhang, Mary ; Chau, David ; Mcmanus, Bruce ; Yang, Decheng. / Specific interaction of HeLa cell proteins with coxsackievirus B3 3′UTR : La autoantigen binds the 3′ and 5′UTR independently of the poly(A) tail. In: Cellular Microbiology. 2007 ; Vol. 9, No. 7. pp. 1705-1715.
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