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Cdc42 GTPase activating proteins Rga4 and Rga6 coordinate septum synthesis and membrane trafficking at the division plane during cytokinesis

Bethany F. Campbell, Brian S. Hercyk, Ashlei R. Williams, Ema San Miguel, Haylee G. Young, View ORCID ProfileMaitreyi E. Das
doi: https://doi.org/10.1101/2021.12.14.472679
Bethany F. Campbell
Department of Biochemistry & Cellular and Molecular Biology, University of Tennessee, Knoxville, TN, USA, 37996
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Brian S. Hercyk
Department of Biochemistry & Cellular and Molecular Biology, University of Tennessee, Knoxville, TN, USA, 37996
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Ashlei R. Williams
Department of Biochemistry & Cellular and Molecular Biology, University of Tennessee, Knoxville, TN, USA, 37996
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Ema San Miguel
Department of Biochemistry & Cellular and Molecular Biology, University of Tennessee, Knoxville, TN, USA, 37996
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Haylee G. Young
Department of Biochemistry & Cellular and Molecular Biology, University of Tennessee, Knoxville, TN, USA, 37996
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Maitreyi E. Das
Department of Biochemistry & Cellular and Molecular Biology, University of Tennessee, Knoxville, TN, USA, 37996
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  • ORCID record for Maitreyi E. Das
  • For correspondence: mdas@utk.edu
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Abstract

Fission yeast cytokinesis is driven by simultaneous septum synthesis, membrane furrowing and actomyosin ring constriction. The septum consists of a primary septum flanked by secondary septa. First, delivery of the glucan synthase Bgs1 and membrane vesicles initiate primary septum synthesis and furrowing. Next, Bgs4 is delivered for secondary septum formation. It is unclear how septum synthesis is coordinated with membrane furrowing. Cdc42 promotes delivery of Bgs1 but not Bgs4. We find that after primary septum initiation, Cdc42 inactivators Rga4 and Rga6 localize to the division site. In rga4Δrga6Δ mutants Cdc42 activity is enhanced during late cytokinesis and cells take longer to separate. Electron micrographs of the division site in these mutants exhibit malformed septum with irregular membrane structures. These mutants have a larger division plane with enhanced Bgs1 delivery but fail to enhance accumulation of Bgs4 and several exocytic proteins. Additionally, these mutants show endocytic defects at the division site. This suggests that Cdc42 regulates only specific membrane trafficking events. Our data indicate that while active Cdc42 promotes primary septum synthesis, as cytokinesis progresses Rga4 and Rga6 localize to the division site to decrease Cdc42 activity. This couples specific membrane trafficking events with septum formation to allow proper septum morphology.

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Synopsis The GTPase Cdc42 regulates cytokinesis in cell-walled fission yeast. Active Cdc42 promotes the initiation of septum (new cell wall) synthesis to physically divide daughter cells. Here we show that Cdc42 activity must be decreased at the later stages of cytokinesis to enable proper septum formation. Mutants lacking Cdc42 inactivators, Rga4 and Rga6, lead to membrane trafficking defects and malformed septa consequently delaying cell separation.

Competing Interest Statement

The authors have declared no competing interest.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted December 14, 2021.
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Cdc42 GTPase activating proteins Rga4 and Rga6 coordinate septum synthesis and membrane trafficking at the division plane during cytokinesis
Bethany F. Campbell, Brian S. Hercyk, Ashlei R. Williams, Ema San Miguel, Haylee G. Young, Maitreyi E. Das
bioRxiv 2021.12.14.472679; doi: https://doi.org/10.1101/2021.12.14.472679
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Cdc42 GTPase activating proteins Rga4 and Rga6 coordinate septum synthesis and membrane trafficking at the division plane during cytokinesis
Bethany F. Campbell, Brian S. Hercyk, Ashlei R. Williams, Ema San Miguel, Haylee G. Young, Maitreyi E. Das
bioRxiv 2021.12.14.472679; doi: https://doi.org/10.1101/2021.12.14.472679

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