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PMID: 10757782 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Domains in the SPT5 protein that modulate its transcriptional regulatory properties.

Molecular and cellular biology ·Vol. 20 ·No. 9 ·2000-05-00 ·Pages 2970-83

Ivanov D, Kwak YT, Guo J, Gaynor RB

Abstract

SPT5 and its binding partner SPT4 regulate transcriptional elongation by RNA polymerase II. SPT4 and SPT5 are involved in both 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB)-mediated transcriptional inhibition and the activation of transcriptional elongation by the human immunodeficiency virus type 1 (HIV-1) Tat protein. Recent data suggest that P-TEFb, which is composed of CDK9 and cyclin T1, is also critical in regulating transcriptional elongation by SPT4 and SPT5. In this study, we analyze the domains of SPT5 that regulate transcriptional elongation in the presence of either DRB or the HIV-1 Tat protein. We demonstrate that SPT5 domains that bind SPT4 and RNA polymerase II, in addition to a region in the C terminus of SPT5 that contains multiple heptad repeats and is designated CTR1, are critical for in vitro transcriptional repression by DRB and activation by the Tat protein. Furthermore, the SPT5 CTR1 domain is a substrate for P-TEFb phosphorylation. These results suggest that C-terminal repeats in SPT5, like those in the RNA polymerase II C-terminal domain, are sites for P-TEFb phosphorylation and function in modulating its transcriptional elongation properties.

MeSH Terms
Amino Acid Sequence Animals Blotting, Western COS Cells Cell Nucleus/metabolism Chromosomal Proteins, Non-Histone Cyclin-Dependent Kinase 9 Cyclin-Dependent Kinases/metabolism Dichlororibofuranosylbenzimidazole/pharmacology Enzyme Inhibitors/pharmacology Fungal Proteins/chemistry,genetics,metabolism Gene Expression Regulation Gene Products, tat/metabolism HeLa Cells Humans Molecular Sequence Data Mutagenesis Nuclear Proteins/chemistry,genetics,metabolism Phosphorylation Positive Transcriptional Elongation Factor B Precipitin Tests Protein Binding Protein Serine-Threonine Kinases/metabolism Protein Structure, Tertiary RNA Polymerase II/metabolism Recombinant Fusion Proteins/metabolism Saccharomyces cerevisiae Proteins Sequence Homology, Amino Acid Transcription, Genetic Transcriptional Elongation Factors
Chemicals
Chromosomal Proteins, Non-Histone Enzyme Inhibitors Fungal Proteins Gene Products, tat Nuclear Proteins Recombinant Fusion Proteins SPT4 protein, S cerevisiae Saccharomyces cerevisiae Proteins Transcriptional Elongation Factors SPT5 transcriptional elongation factor Dichlororibofuranosylbenzimidazole Positive Transcriptional Elongation Factor B Protein Serine-Threonine Kinases CDK9 protein, human Cyclin-Dependent Kinase 9 Cyclin-Dependent Kinases RNA Polymerase II
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Ivanov D
Division of Hematology-Oncology, Department of Medicine, Harold Simmons Cancer Center, University of Texas Southwestern Medical Center, Dallas, Texas 75235-8594, USA.
Kwak Y T
Guo J
Gaynor R B
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.20.9.2970-2983.2000
Published
2000-05-00
Pages
2970-83
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC85557
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.20.9.2970-2983.2000 PMC Full Text View journal details
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