Rpb4p is necessary for RNA polymerase II activity at high temperature.
Rpb4p and Rpb7p are two subunits of the yeast RNA polymerase II, which form a subcomplex that can dissociate from the enzyme in vitro. Whereas RPB7 is essential, RPB4 is dispensable for cellular viability. However, the rpb4 null mutant is heat-sensitive, and it has been suggested that Rpb4p is an essential component for cellular stress response. To examine this hypothesis, we used two-dimensional gel electrophoresis to analyze the protein expression pattern of the rpb4 null mutant in response to heat shock, oxidative stress, osmotic stress, and in the post-diauxic phase. We show that this mutant is not impaired in stress induced transcriptional activation: the absence of heat shock response of the mutant is due to a general defect in RNA polymerase II activity at high temperature. Under this condition, Rpb4p is necessary to maintain the polymerase activity in vivo. The heat growth defect of the rpb4 null mutant can be partially suppressed by overexpression of RPB7, suggesting that Rpb4p maintains or stabilizes Rpb7p in the RNA polymerase. We also demonstrate that rpb4 null mutant is an appropriate tool to analyze the involvement of transcriptional events in the survival and adaptation to heat shock or other stresses.
Maillet
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Service de Biochimie et Génétique Moléculaire, CEA-Saclay, F-91191 Gif-sur-Yvette Cedex, France.
Buhler
J M
JM
Sentenac
A
A
Labarre
J
J
D015180:Electrophoresis, Gel, Two-Dimensional; D004795:Enzyme Stability; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D018869:Heat-Shock Response / Q000502:physiology*; D006358:Hot Temperature; D009154:Mutation; D009997:Osmotic Pressure; D018384:Oxidative Stress / Q000502:physiology; D012319:RNA Polymerase II / Q000235:genetics / Q000378:metabolism*; D015533:Transcriptional Activation
D005656:Fungal Proteins; D012319:RNA Polymerase II