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PMID: 12604794 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

von Hippel-Lindau protein binds hyperphosphorylated large subunit of RNA polymerase II through a proline hydroxylation motif and targets it for ubiquitination.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 100 ·No. 5 ·2003-03-04 ·Pages 2706-11

Kuznetsova AV, Meller J, Schnell PO, Nash JA, Ignacak ML, Sanchez Y, Conaway JW, Conaway RC, Czyzyk-Krzeska MF

Abstract

The transition from transcription initiation to elongation involves phosphorylation of the large subunit (Rpb1) of RNA polymerase II on the repetitive carboxyl-terminal domain. The elongating hyperphosphorylated Rpb1 is subject to ubiquitination, particularly in response to UV radiation and DNA-damaging agents. By using computer modeling, we identified regions of Rpb1 and the adjacent subunit 6 of RNA polymerase II (Rpb6) that share sequence and structural similarity with the domain of hypoxia-inducible transcription factor 1 alpha (HIF-1 alpha) that binds von Hippel-Lindau tumor suppressor protein (pVHL). pVHL confers substrate specificity to the E3 ligase complex, which ubiquitinates HIF-alpha and targets it for proteasomal degradation. In agreement with the computational model, we show biochemical evidence that pVHL specifically binds the hyperphosphorylated Rpb1 in a proline-hydroxylation-dependent manner, targeting it for ubiquitination. This interaction is regulated by UV radiation.

MeSH Terms
Amino Acid Motifs Amino Acid Sequence Animals Biotinylation Blotting, Western Cell Nucleus/metabolism DNA Damage Ligases/metabolism,physiology Models, Molecular Molecular Sequence Data Oxygen/metabolism PC12 Cells Phosphorylation Precipitin Tests Proline/chemistry Protein Binding Protein Structure, Tertiary RNA Polymerase II/chemistry,metabolism Rats Sequence Homology, Amino Acid Software Substrate Specificity Tumor Suppressor Proteins Ubiquitin/metabolism Ubiquitin-Protein Ligases Ultraviolet Rays Von Hippel-Lindau Tumor Suppressor Protein
Chemicals
Tumor Suppressor Proteins Ubiquitin Proline Ubiquitin-Protein Ligases Von Hippel-Lindau Tumor Suppressor Protein RNA Polymerase II Ligases Oxygen
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Kuznetsova Anna V
Department of Molecular and Cellular Physiology, Children's Hospital Research Foundation, University of Cincinnati College of Medicine, Cincinnati, OH 45267-0576, USA.
Meller Jaroslaw
Schnell Phillip O
Nash James A
Ignacak Monika L
Sanchez Yolanda
Conaway Joan W
Conaway Ronald C
Czyzyk-Krzeska Maria F
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.0436037100
Published
2003-03-04
Epub
2003-00-25
Pages
2706-11
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC151405
Subset
IM
Grants
NHLBI NIH HHS · R01 HL058687 · United States
NHLBI NIH HHS · R01 HL066312 · United States
NHLBI NIH HHS · HL66312 · United States
PHS HHS · L58687 · United States
External Links
PubMed source DOI: 10.1073/pnas.0436037100 PMC Full Text View journal details
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