3 RNA Polymerase II Subunits, Transcription Factors, and Kinases: Investigations in Drosophila melanogaster and Saccharomyces cerevisiae

Abstract

OVERVIEW
An approach combining detailed biochemistry with classic and molecular genetics has led to new insights into the structure and function of RNA polymerase II (pol II), its transcription factors, and a novel RNA pol II kinase. Collections of mutants altered in genes for the two large subunits of RNA pol II will be valuable for elucidating the functions of conserved domains discovered as a consequence of cloning and sequencing the genes. A fractionated, RNA pol II-dependent transcription system should continue to contribute to the functional analysis of mutant enzymes as well as to characterization of transcription factors and their interactions with the polymerase. Investigations of a fascinating protein kinase that phosphorylates the unusual carboxy-terminal repeat domain of subunit IIa should generate new insights into novel regulatory phenomena involving RNA pol II.

INTRODUCTION
We have been investigating the eukaryotic transcription machinery, focusing on RNA pol II and factors with which it interacts. Our rationale has been that in order to understand mechanisms regulating class II gene transcription, we need to know in some detail the structural and functional features of RNA pol II, especially those that concern its interactions with general and regulatory transcription factors. Although our work has been primarily with the fruit fly Drosophila melanogaster, the structural conservation of the transcription apparatus during evolution suggests that most of our results will be applicable to eukaryotes in general. Our approach to characterizing RNA pol II has been to combine genetics with biochemistry. In addition, when feasible, we have persisted...