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Table II.

The GAP activity of Msb3p is required for the suppression of the actin-organization defects incdc42-Ts mutants


Plasmids

YEF115 (cdc42-1) host strain

YEF2258 (cdc42-201) host strain
 (24°C) 0 min  38.5°C 5.3 h  (24°C) 0 min  35.5°C 6 h  

 
Budded
 cells
 
Actin
 polarized
 
Budded
 cells
 
Actin
 polarized
 
Budded
 cells
 
Actin
 polarized
 
Budded
 cells
 
Actin
 polarized
 

 
%
 
%
 
%
 
%
 
%
 
%
 
%
 
%
 
YEplac181 20 
YEp181-3HA-MSB3 42 75 48 38 
YEp181-3HA-MSB3-R282K 10 13 39 
YEp181-3HA-MSB1
 
4
 
6
 
29
 
60
 
7
 
3
 
20
 
19
 

Plasmids

YEF115 (cdc42-1) host strain

YEF2258 (cdc42-201) host strain
 (24°C) 0 min  38.5°C 5.3 h  (24°C) 0 min  35.5°C 6 h  

 
Budded
 cells
 
Actin
 polarized
 
Budded
 cells
 
Actin
 polarized
 
Budded
 cells
 
Actin
 polarized
 
Budded
 cells
 
Actin
 polarized
 

 
%
 
%
 
%
 
%
 
%
 
%
 
%
 
%
 
YEplac181 20 
YEp181-3HA-MSB3 42 75 48 38 
YEp181-3HA-MSB3-R282K 10 13 39 
YEp181-3HA-MSB1
 
4
 
6
 
29
 
60
 
7
 
3
 
20
 
19
 

The enriched, unbudded population of cells harboring indicated plasmids were grown in fresh media at indicated temperatures for the indicated lengths of time before assaying for the suppression of the budding and the actin-organization defects (see Materials and methods for detail). More than 600 cells were scored to determine the percentage of budded cells by DIC microscopy, and at least 200 cells were scored for actin organization through F-actin staining. Shown here is one set of representative results from three independent experiments performed in each cdc42-Ts host strain. Representative cells are shown in Fig. 5 B.

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