Calcium-dependent inactivation of RNA polymerase III transcription.
Extracts from whole oocytes of Xenopus laevis are widely used as an efficient in vitro system for the transcription of cloned genes by RNA polymerase III. We have found that these extracts no longer support RNA polymerase III transcription in response to a brief incubation in the presence of Ca2+. However, when transcription complexes were first formed on the genes, a subsequent incubation in the presence of Ca2+ had little effect. Fractionation of extracts was used to show that transcription factors (TF) IIIC and, to a lesser extent, TFIIIB, but not RNA polymerase III, were targets of the Ca(2+)-dependent inactivation process. An additional component (not present in fractionated TFIIIC or TFIIIB) was required for the Ca(2+)-dependent destruction of transcription factor activity. The Ca(2+)-dependent inactivation process was blocked by protease inhibitors that inhibit known Ca(2+)-dependent proteases called calpains. These results suggest that TFIIIC and TFIIIB are inactivated by an endogenous calpain. The common use of Ca2+ as a second messenger and the widespread distribution of calpains suggest that the proteolytic degradation of transcription factors may be a general mechanism for the regulation of gene expression.
Sturges
M R
MR
Sinsheimer Laboratories, University of California, Santa Cruz 95064.
Peck
L J
LJ
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D011480:Protease Inhibitors / Q000494:pharmacology; D012320:RNA Polymerase III / Q000378:metabolism*; D012341:RNA, Ribosomal, 5S / Q000235:genetics; D035361:Transcription Factor TFIIIB; D014157:Transcription Factors / Q000037:antagonists & inhibitors / Q000378:metabolism; D019474:Transcription Factors, TFIII; D014158:Transcription, Genetic / Q000187:drug effects; D014982:Xenopus laevis; D000098832:Transcription Factor TFIIIC
D011480:Protease Inhibitors; D012341:RNA, Ribosomal, 5S; D035361:Transcription Factor TFIIIB; D014157:Transcription Factors; D019474:Transcription Factors, TFIII; D000098832:Transcription Factor TFIIIC; D012320:RNA Polymerase III; D002118:Calcium