TAILIEUCHUNG - Lecture Molecular biology: Chapter 6 - Robert F. Weaver

Chapter 6 - The mechanism of transcription in bacteria. In this chapter we will focus on the basic mechanism of transcription. We will begin with RNA polymerase, the enzyme that catalyzes transcription. We will also look at the interaction between RNA polymerase and DNA. | Molecular Biology Fourth Edition Chapter 6 The Mechanism of Transcription in Bacteria Lecture PowerPoint to accompany Robert F. Weaver Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. RNA Polymerase Structure By 1969 SDS-PAGE of RNA polymerase from E. coli had shown several subunits 2 very large subunits are b (150 kD) and b’ (160 kD) Sigma (s) at 70 kD Alpha (a) at 40 kD – 2 copies present in holoenzyme Omega (w) at 10 kD Was not clearly visible in SDS-PAGE, but seen in other experiments Not required for cell viability or in vivo enzyme activity Appears to play a role in enzyme assembly 6- Sigma as a Specificity Factor Core enzyme without the s subunit could not transcribe viral DNA, yet had no problems with highly nicked calf thymus DNA With s subunit, the holoenzyme worked equally well on both types of DNA 6- Testing Transcription Core enzyme transcribes both DNA strands Without s-subunit the core enzyme has basic transcribing

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