Affordable Access

deepdyve-link
Publisher Website

Functions of the TFIIE-Related Tandem Winged-Helix Domain of Rpc34 in RNA Polymerase III Initiation and Elongation.

Authors
  • Wei, Yi-Yu1
  • Chen, Hung-Ta2, 3
  • 1 Graduate Institute of Life Sciences, National Defense Medical Center, Taipei, Taiwan, Republic of China. , (China)
  • 2 Graduate Institute of Life Sciences, National Defense Medical Center, Taipei, Taiwan, Republic of China [email protected] , (China)
  • 3 Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan, Republic of China. , (China)
Type
Published Article
Journal
Molecular and Cellular Biology
Publisher
American Society for Microbiology
Publication Date
Feb 15, 2018
Volume
38
Issue
4
Identifiers
DOI: 10.1128/MCB.00105-17
PMID: 29180511
Source
Medline
Keywords
License
Unknown

Abstract

Rpc34 is a subunit of the Rpc82/34/31 subcomplex residing on the DNA-binding cleft of RNA polymerase (Pol) III. Rpc34 contains a structurally flexible N-terminal tandem winged-helix (tWH) domain related to the TFIIE transcription factor. While the second WH (WH2) fold of the tWH domain is known to function in DNA melting activity during transcription initiation, the functional role of the WH1 fold is unknown. In this study, we generated a series of new Rpc34 tWH mutants conferring a cold-sensitive growth phenotype. We found that the tWH mutations severely compromised in vitro transcription activity due to destabilization of the preinitiation complex (PIC). Site-specific protein photo-cross-linking analysis indicated that the tWH domain persistently interacts with protein subunits of the Pol III cleft in the PIC and the ternary elongation complex (TEC). Furthermore, purified Pol III proteins with tWH mutations also showed reduced efficiency in RNA elongation. Our study results suggest that the tWH domain is an important protein module above the Pol III cleft that integrates protein and nucleic acid interactions for initiation and elongation.

Report this publication

Statistics

Seen <100 times