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Involvement of Vts1, a structure-specific RNA-binding protein, in Okazaki fragment processing in yeast

Cited 9 time in Web of Science Cited 9 time in Scopus
Authors

Lee, Chul-Hwan; Shin, Yong-Keol; Phung, Thi Thu Huong; Bae, Jae Seok; Kang, Young-Hoon; Nguyen, Tuan Anh; Kim, Jeong-Hoon; Kim, Do-Hyung; Kang, Min-Jung; Bae, Sung-Ho; Seo, Yeon-Soo

Issue Date
2010-03
Publisher
Oxford University Press
Citation
Nucleic Acids Research, Vol.38 No.5, pp.1583-1595
Abstract
The non-essential VTS1 gene of Saccharomyces cerevisiae is highly conserved in eukaryotes and encodes a sequence- and structure-specific RNA-binding protein. The Vts1 protein has been implicated in post-transcriptional regulation of a specific set of mRNAs that contains its-binding site at their 30-untranslated region. In this study, we identified VTS1 as a multi-copy suppressor of dna2-K1080E, a lethal mutant allele of DNA2 that lacks DNA helicase activity. The suppression was allele-specific, since overexpression of Vts1 did not suppress the temperature-dependent growth defects of dna2Δ405N devoid of the N-terminal 405-amino-acid residues. Purified recombinant Vts1 stimulated the endonuclease activity of wild-type Dna2, but not the endonuclease activity of Dna2"405N, indicating that the activation requires the N-terminal domain of Dna2. Stimulation of Dna2 endonuclease activity by Vts1 appeared to be the direct cause of suppression, since the multi-copy expression of Dna2-K1080E suppressed the lethality observed with its singlecopy expression. We found that vts1Δ dna2Δ405N and vts1Δdna2-7 double mutant cells displayed synergistic growth defects, in support of a functional interaction between two genes. Our results provide both in vivo and in vitro evidence that Vts1 is involved in lagging strand synthesis by modulating the Dna2 endonuclease activity that plays an essential role in Okazaki fragment processing. © The Author(s) 2009. Published by Oxford University Press.
ISSN
0305-1048
URI
https://hdl.handle.net/10371/201860
DOI
https://doi.org/10.1093/nar/gkp1135
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