Analysis of the RNA binding specificity of the human tap protein, a constitutive transport element-specific nuclear RNA export factor

Yibin Kang, Hal P. Bogerd, Jin Yang, Bryan R. Cullen

Research output: Contribution to journalArticlepeer-review

33 Scopus citations

Abstract

The human Tap protein has been proposed to mediate Mason Pfizer monkey virus constitutive transport element (CTE)-dependent nuclear RNA export and may also play a role in global mRNA export. Here, we have used in vivo assays, in both yeast and human cells, together with in vitro assays, to further characterize the RNA binding properties of Tap, which has been proposed to contain a novel leucine-rich RNA binding motif. Using the yeast three hybrid assay, we selected RNA molecules that retain Tap binding activity from a pool of randomized CTE sequences. The recovered RNA sequences differed only minimally from the wild-type CTE yet all displayed lower affinity for Tap both in vivo and in vitro. Analysis of the RNA export activity of the recovered CTE variants revealed that Tap affinity was highly predictive of CTE biological activity. Together, these observations provide additional evidence supporting the identification of Tap as the direct cofactor for CTE function and demonstrate that RNA binding by Tap is highly sequence specific.

Original languageEnglish (US)
Pages (from-to)200-209
Number of pages10
JournalVirology
Volume262
Issue number1
DOIs
StatePublished - Sep 15 1999
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Virology

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