Lte1 contributes to Bfa1 localization rather than stimulating nucleotide exchange by Tem1.

The Journal of Cell Biology
Marco GeymonatSteven G Sedgwick

Abstract

Lte1 is a mitotic regulator long envisaged as a guanosine nucleotide exchange factor (GEF) for Tem1, the small guanosine triphosphatase governing activity of the Saccharomyces cerevisiae mitotic exit network. We demonstrate that this model requires reevaluation. No GEF activity was detectable in vitro, and mutational analysis of Lte1's putative GEF domain indicated that Lte1 activity relies on interaction with Ras for localization at the bud cortex rather than providing nucleotide exchange. Instead, we found that Lte1 can determine the subcellular localization of Bfa1 at spindle pole bodies (SPBs). Under conditions in which Lte1 is essential, Lte1 promoted the loss of Bfa1 from the maternal SPB. Moreover, in cells with a misaligned spindle, mislocalization of Lte1 in the mother cell promoted loss of Bfa1 from one SPB and allowed bypass of the spindle position checkpoint. We observed that lte1 mutants display aberrant localization of the polarity cap, which is the organizer of the actin cytoskeleton. We propose that Lte1's role in cell polarization underlies its contribution to mitotic regulation.

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Citations

Jun 29, 2011·Proceedings of the National Academy of Sciences of the United States of America·Jill E FalkAngelika Amon
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Dec 15, 2010·The Journal of Cell Biology·Marco GeymonatSteven G Sedgwick
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Jul 1, 2020·Cell Discovery·Roberto QuadriMarco Muzi-Falconi
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Nov 13, 2020·Frontiers in Cell and Developmental Biology·Roberto QuadriMarco Muzi-Falconi
Jun 24, 2021·Cell Discovery·Roberto QuadriMarco Muzi-Falconi

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Methods Mentioned

BETA
GTPase
nucleotide exchange
PCR
affinity purification
fluorescence microscopy

Software Mentioned

Photoshop CS
SoftWoRx
ImageJ

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