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| Status |
Public on Jan 01, 2013 |
| Title |
Pronounced and Extensive Microtubule Defects in a Saccharomyces cerevisiae DIS3 Mutant |
| Organism |
Saccharomyces cerevisiae |
| Experiment type |
Expression profiling by high throughput sequencing
|
| Summary |
The recently proposed exozyme hypothesis posits that subunits of the RNA processing exosome assemble into structurally distinct protein complexes that function in disparate cellular compartments and RNA metabolic pathways. Here, in a genetic test of this hypothesis, we examine the role of Dis3 -- an essential polypeptide with endo- and 3' to 5' exo-ribonuclease activity -- in cell cycle progression. We present several lines of evidence that perturbation of DIS3 affects microtubule (MT) localization and structure in Saccharomyces cerevisiae. Cells with a DIS3 mutation: (i) accumulate anaphase and pre-anaphase mitotic spindles; (ii) exhibit spindles that are mis-oriented and displaced from the bud neck; (iii) harbor elongated spindle-associated astral MTs; (iv) have an increased G1 astral MT length and number; and (v) are hypersensitive to MT poisons. Mutations in the core exosome genes RRP4 and MTR3 and the exosome cofactor gene MTR4 -- but not other exosome subunit gene mutants -- also elicit MT phenotypes. RNA deep sequencing analysis (RNA-seq) shows broad changes in the levels of cell cycle- and microtubule-related transcripts in mutant strains. Collectively, the different mitotic phenotypes and distinct sets of mRNAs affected by the exosome subunit and cofactor mutants studied here suggest that Dis3 has a core exosome-independent role(s) in cell cycle progression. These observations are consistent with the predictions of the exozyme hypothesis and also suggest an evolutionarily conserved role for Dis3 in linking RNA metabolism, MTs, and mitotic progression.
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| Overall design |
RNA-seq analysis of total RNA harvested from WT, mtr3-1, mtr4-1, and Dis3^mtr (rrp44-1/mtr17-1) Saccharomyces cerevisiae strains after a temperature shift.
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| Contributor(s) |
Kiss DL, Andrulis ED |
| Citation(s) |
21919057 |
| |
| Submission date |
May 02, 2011 |
| Last update date |
May 15, 2019 |
| Contact name |
Daniel Louis Kiss |
| E-mail(s) |
dlk13@case.edu
|
| Phone |
216-368-0385
|
| Fax |
216-368-3055
|
| Organization name |
Case Western Reserve University
|
| Department |
Molceular Biology and Microbiology
|
| Lab |
Andrulis
|
| Street address |
2109 Adelbert Road
|
| City |
Cleveland |
| State/province |
OH |
| ZIP/Postal code |
44106 |
| Country |
USA |
| |
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| Platforms (1) |
| GPL13272 |
Illumina Genome Analyzer IIx (Saccharomyces cerevisiae) |
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| Samples (4)
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| Relations |
| SRA |
SRP006672 |
| BioProject |
PRJNA140473 |
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