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PMID: 18285459 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

The von Hippel-Lindau tumor suppressor protein and Egl-9-Type proline hydroxylases regulate the large subunit of RNA polymerase II in response to oxidative stress.

Molecular and cellular biology ·Vol. 28 ·No. 8 ·2008-04-00 ·Pages 2701-17

Mikhaylova O, Ignacak ML, Barankiewicz TJ, Harbaugh SV, Yi Y, Maxwell PH, Schneider M, Van Geyte K, Carmeliet P, Revelo MP, Wyder M, Greis KD, Meller J, Czyzyk-Krzeska MF

Abstract

Human renal clear cell carcinoma (RCC) is frequently associated with loss of the von Hippel-Lindau (VHL) tumor suppressor (pVHL), which inhibits ubiquitylation and degradation of the alpha subunits of hypoxia-inducible transcription factor. pVHL also ubiquitylates the large subunit of RNA polymerase II, Rpb1, phosphorylated on serine 5 (Ser5) within the C-terminal domain (CTD). A hydroxylated proline 1465 within an LXXLAP motif located N-terminal to the CTD allows the interaction of Rpb1 with pVHL. Here we report that in RCC cells, pVHL regulates expression of Rpb1 and is necessary for low-grade oxidative-stress-induced recruitment of Rpb1 to the DNA-engaged fraction and for its P1465 hydroxylation, phosphorylation, and nondegradative ubiquitylation. Egln-9-type prolyl hydroxylases, PHD1 and PHD2, coimmunoprecipitated with Rpb1 in the chromatin fraction of VHL(+) RCC cells in response to oxidative stress, and PHD1 was necessary for P1465 hydroxylation while PHD2 had an inhibitory effect. P1465 hydroxylation was required for oxidative-stress-induced Ser5 phosphorylation of Rpb1. Importantly, overexpression of wild-type Rpb1 stimulated formation of kidney tumors by VHL(+) cells, and this effect was abolished by P1465A mutation of Rpb1. These data indicate that through this novel pathway involving P1465 hydroxylation and Ser5 phosphorylation of Rbp1, pVHL may regulate tumor growth.

MeSH Terms
Animals Cell Line DNA-Binding Proteins/deficiency,genetics,metabolism Humans Hypoxia-Inducible Factor-Proline Dioxygenases Immediate-Early Proteins/deficiency,genetics,metabolism Mice Mice, Knockout Oxidative Stress Phosphoserine/metabolism Procollagen-Proline Dioxygenase/deficiency,genetics,metabolism Protein Binding Protein Subunits/genetics,metabolism RNA Polymerase II/metabolism Retinol-Binding Proteins, Cellular/metabolism Ubiquitination Von Hippel-Lindau Tumor Suppressor Protein/genetics,metabolism
Chemicals
DNA-Binding Proteins Immediate-Early Proteins Protein Subunits RBP1 protein, human Retinol-Binding Proteins, Cellular Phosphoserine PHD1 protein, mouse Procollagen-Proline Dioxygenase Egln1 protein, mouse Hypoxia-Inducible Factor-Proline Dioxygenases Von Hippel-Lindau Tumor Suppressor Protein RNA Polymerase II
Authors & Affiliations
14 authors, click to expand affiliations / ORCID
Mikhaylova Olga
Department of Molecular Oncogenesis, Genome Research Institute, University of Cincinnati, Cincinnati, OH 45237-0505, USA. maria.czyzykkrzeska@uc.edu
Ignacak Monika L
Barankiewicz Teresa J
Harbaugh Svetlana V
Yi Ying
Maxwell Patrick H
Schneider Martin
Van Geyte Katie
Carmeliet Peter
Revelo Monica P
Wyder Michael
Greis Kenneth D
Meller Jarek
Czyzyk-Krzeska Maria F
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
1098-5549
DOI
10.1128/MCB.01231-07
Email
maria.czyzykkrzeska@uc.edu
Published
2008-04-00
Epub
2008-00-19
Pages
2701-17
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC2293119
Subset
IM
Grants
NHLBI NIH HHS · R01 HL058687 · United States
British Heart Foundation · United Kingdom
NHLBI NIH HHS · HL58687 · United States
NCI NIH HHS · R01 CA122346 · United States
NCI NIH HHS · CA122346 · United States
NHLBI NIH HHS · HL66312 · United States
NHLBI NIH HHS · R01 HL066312 · United States
External Links
PubMed source DOI: 10.1128/MCB.01231-07 PMC Full Text View journal details
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