GTP hydrolysis is not important for Ypt1 GTPase function in vesicular transport.

Molecular and Cellular Biology
C J RichardsonN Segev

Abstract

GTPases of the Ypt/Rab family play a key role in the regulation of vesicular transport. Their ability to cycle between the GTP- and the GDP-bound forms is thought to be crucial for their function. Conversion from the GTP- to the GDP-bound form is achieved by a weak endogenous GTPase activity, which can be stimulated by a GTPase-activating protein (GAP). Current models suggest that GTP hydrolysis and GAP activity are essential for vesicle fusion with the acceptor compartment or for timing membrane fusion. To test this idea, we inactivated the GTPase activity of Ypt1p by using the Q67L mutation, which targets a conserved residue that helps catalyze GTP hydrolysis in Ras. We demonstrate that the mutant Ypt1-Q67L protein is severely impaired in its ability to hydrolyze GTP both in the absence and in the presence of GAP and consequently is restricted mostly to the GTP-bound form. Surprisingly, a strain with ypt1-Q67L as the only YPT1 gene in the cell has no observable growth phenotypes at temperatures ranging from 14 to 37 degrees C. In addition, these mutant cells exhibit normal rates of secretion and normal membrane morphology as determined by electron microscopy. Furthermore, the ypt1-Q67L allele does not exhibit dominant phenoty...Continue Reading

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Citations

Jul 24, 2013·The Journal of Cell Biology·Francis A Barr
Aug 19, 2008·Trends in Cell Biology·Hao WuPatrick Brennwald
Sep 20, 2008·Traffic·Michael SacherNava Segev
Oct 15, 2014·British Journal of Haematology·MinJung KimCurt I Civin
Aug 17, 1999·Current Opinion in Cell Biology·P Chavrier, B Goud
Jul 27, 2000·Molecular Cell·M HetzerI W Mattaj
Oct 2, 2001·Science's STKE : Signal Transduction Knowledge Environment·N Segev
Oct 8, 1998·Molecular Biology of the Cell·S JonesN Segev
Apr 24, 2009·Molecular Biology of the Cell·Kimberly Romero RosalesAimee L Edinger
Aug 9, 2005·Nature Cell Biology·Alexander K HaasFrancis A Barr
Feb 24, 2015·Critical Reviews in Biochemistry and Molecular Biology·Zhanna LipatovaNava Segev
Jul 15, 2015·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Chang Ho KangSang Yeol Lee
Jan 12, 2001·Physiological Reviews·Y TakaiT Matozaki
Jul 17, 2001·Current Opinion in Cell Biology·N Segev
Aug 6, 2008·Plant Physiology·Enrique Rojo, Jurgen Denecke

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