A genetic screen in Saccharomyces cerevisiae identifies new genes that interact with mex67-5, a temperature-sensitive allele of the gene encoding the mRNA export receptor
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The Mex67p protein, together with Mtr2p, functions as the mRNA export receptor in Saccharomyces cerevisiae by interacting with both mRNA and nuclear pore complexes. To identify genes that interact functionally with MEX67, we used transposon insertion to search for mutations that suppressed the temperature-sensitive mex67-5 allele. Four suppressors are described here. The screen revealed that mutant Mex67-5p, but not wild-type Mex67p, is a target of the nuclear protein quality control mediated by San1p, a ubiquitin-protein ligase that participates in degradation of aberrant chromatin-associated proteins. Our finding that overexpression of the SPT6 gene alleviates the growth defects of the mex67-5 strain, together with the impairment of poly(A)+ RNA export caused by depletion of Spt6p or the related protein Iws1p/Spn1p, supports the mechanism proposed in mammalian cells for Spt6-mediated co-transcriptional loading of mRNA export factors during transcription elongation. Finally, our results also uncovered genetic connections between Mex67p and the poly(A) nuclease complex and with components of chromatin boundary elements.
KeywordsmRNA export Mex67p Transcription Protein quality control Barrier complex Poly(A) nuclease
We thank S. Chávez, S. Rodríguez-Navarro and M. Pamblanco for providing plasmids and yeast strains. This work was supported by grants from the Spanish Ministry of Education and Science (BFU2004-00069/BMC and BFU2007-61525/BMC) to F.E and from the National Institutes of Health (GM33998) to C.N.C. L.P.-C. is recipient of a fellowship from the F.P.I program of the Spanish Ministry of Education and Science.
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