Identification and characterization of supressors for rng15 deletion in fission yeast cytokinesis
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Date
2016-03-30
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Abstract
While much is known about cytokinesis, the final stage of
cell-division cycle, our knowledge on roles of vesicle
trafficking during cytokinesis is limited. To further our
understanding of exocytosis during cytokinesis, we
investigate Rng15 in fission yeast, a popular model organism
for cytokinesis. Little is known about Rng15, though our
preliminary studies and its homology with budding yeast
Mso1 suggest that it participates in exocytosis. Rng15 also
shares homology to Mint1, a mammalian adaptor protein
involved in exocytosis. Cells with rng15 gene deleted (rng15Δ)
cannot grow at 36°C due to defective cytokinesis and
accumulate secretory vesicles at the division site.
To further investigate how Rng15 functions in cytokinesis,
we screened for high-copy suppressors of rng15Δ at 36°C by
transforming rng15Δ cells with plasmid DNA from a fission
yeast genomic DNA library. We reasoned that if the proteins
being produced from the plasmids rescue the growth of rng15
Δ cells at 36°C, these proteins are involved in similar
pathways as Rng15. So far we have isolated that Rng15 and
Gmh5 from the screen. Gmh5 is a membrane protein of the
Golgi mannosyltransferase complex, which is predicted to be
involved in elongation of the polysaccharide mannan
backbone and cell wall biogenesis. We will study Gmh5
functions by testing its localization, deletion phenotype, and
genetic interactions with rng15Δ and other mutations in
cytokinesis. In addition, we plan to identify more suppressors
with additional screen. Through this investigation, we will
have a better understanding of the role of exocytosis in the
delivery of materials during fission yeast cytokinesis.
Furthermore, we hope to provide others in the field with
potential insights on how Mso1/Rng15 family proteins work in
cytokinesis in other model systems, such as mammals.
Description
Biological Sciences (The Ohio State University Denman Undergraduate Research Forum)
Keywords
S. pombe, cytokinesis, rng15, gmh5