Hammell, C. M., Gross, S., Zenklusen, D., Heath, C. V., Stutz, F., Moore, C., Cole, C. N. (2002) Coupling of termination, 3′ processing, and mRNA export. Molecular and Cellular Biology, 22 (18). pp. 6441-6457. ISSN 02707306 (ISSN)
Abstract
In a screen to identify genes required for mRNA export in Saccharomyces cerevisiae, we isolated an allele of poly(A) polymerase (PAP1) and novel alleles encoding several other 3′ processing factors. Many newly isolated and some previously described mutants (rna14-48, rna14-49, rna14-64, rna15-58, and pcf11-1 strains) are defective in polymerase II (Pol II) termination but, interestingly, retain the ability to polyadenylate these improperly processed transcripts at the nonpermissive temperature. Deletion of the cis-acting sequences required to couple 3′ processing and termination also produces transcripts that fail to exit the nucleus, suggesting that all of these processes (cleavage, termination, and export) are coupled. We also find that several but not all mRNA export mutants produce improperly 3′ processed transcripts at the nonpermissive temperature. 3′ maturation defects in mRNA export mutants include improper Pol II termination and/or the previously characterized hyperpolyadenylation of transcripts. Importantly, not all mRNA export mutants have defects in 3′ processing. The similarity of the phenotypes of some mRNA export mutants and 3′ processing mutants indicates that some factors from each process may mechanistically interact to couple mRNA processing and export. Consistent with this assumption, we present evidence that Xpo1p interacts in vivo with several 3′ processing factors and that the addition of recombinant Xpo1p to in vitro processing reaction mixtures stimulates 3′ maturation. Of the core 3′ processing factors tested (Rna14p, Rna15p, Pcf11p, Hrp1p, Fip1p, and Cft1p), only Hrp1p shuttles. Overexpression of Rat8p/Dbp5p suppresses both 3′ processing and mRNA export defects found in xpo1-1 cells.
| Item Type: | Paper |
|---|---|
| Uncontrolled Keywords: | cis acting element galactose messenger RNA polynucleotide adenylyltransferase raffinose RNA polymerase II 3' untranslated region allele article cell nucleus controlled study flow cytometry nonhuman polyadenylation priority journal RNA cleavage RNA processing RNA transcription RNA transport Saccharomyces cerevisiae temperature sensitivity two hybrid system Alleles Biological Transport Fungal Proteins Gene Deletion Green Fluorescent Proteins Karyopherins Luminescent Proteins mRNA Cleavage and Polyadenylation Factors Mutation Nuclear Proteins Open Reading Frames Receptors, Cytoplasmic and Nuclear Recombination, Genetic RNA RNA, Messenger RNA-Binding Proteins Saccharomyces cerevisiae Proteins Temperature Transcription, Genetic Two-Hybrid System Techniques Saccharomyces |
| Subjects: | bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification bioinformatics > genomics and proteomics > genetics & nucleic acid processing bioinformatics > genomics and proteomics bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > mRNA dynamics bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > pre-mRNA organism description > yeast |
| CSHL Authors: | |
| Communities: | CSHL labs > Hammell C. lab |
| Depositing User: | Matt Covey |
| Date: | 2002 |
| Date Deposited: | 11 Mar 2013 16:53 |
| Last Modified: | 11 Mar 2013 16:53 |
| PMCID: | PMC135649 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/27739 |
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