Rsp5 Ubiquitin-Protein Ligase Mediates DNA Damage-Induced Degradation of the Large Subunit of RNA Polymerase II in Saccharomyces cerevisiae

ABSTRACT Rsp5 is an E3 ubiquitin-protein ligase of Saccharomyces cerevisiae that belongs to the hect domain family of E3 proteins. We have previously shown that Rsp5 binds and ubiquitinates the largest subunit of RNA polymerase II, Rpb1, in vitro. We show here that Rpb1 ubiquitination and degradation are induced in vivo by UV irradiation and by the UV-mimetic compound 4-nitroquinoline-1-oxide (4-NQO) and that a functional RSP5 gene product is required for this effect. The 26S proteasome is also required; a mutation ofSEN3/RPN2 (sen3-1), which encodes an essential regulatory subunit of the 26S proteasome, partially blocks 4-NQO-induced degradation of Rpb1. These results suggest that Rsp5-mediated ubiquitination and degradation of Rpb1 are components of the response to DNA damage. A human WW domain-containing hect (WW-hect) E3 protein closely related to Rsp5, Rpf1/hNedd4, also binds and ubiquitinates both yeast and human Rpb1 in vitro, suggesting that Rpf1 and/or another WW-hect E3 protein mediates UV-induced degradation of the large subunit of polymerase II in human cells.

[1]  J. Kato,et al.  Degradation of the cyclin-dependent-kinase inhibitor p27Kip1 is instigated by Jab1 , 1999, Nature.

[2]  Xiao Zhen Zhou,et al.  Function of WW domains as phosphoserine- or phosphothreonine-binding modules. , 1999, Science.

[3]  J. Huibregtse,et al.  Functional Domains of the Rsp5 Ubiquitin-Protein Ligase , 1999, Molecular and Cellular Biology.

[4]  Fred Winston,et al.  Functional Organization of the Yeast SAGA Complex: Distinct Components Involved in Structural Integrity, Nucleosome Acetylation, and TATA-Binding Protein Interaction , 1999, Molecular and Cellular Biology.

[5]  C. Michels,et al.  The Role of Ubiquitin Conjugation in Glucose-induced Proteolysis of Saccharomyces Maltose Permease* , 1998, The Journal of Biological Chemistry.

[6]  F E Cohen,et al.  Exploiting the basis of proline recognition by SH3 and WW domains: design of N-substituted inhibitors. , 1998, Science.

[7]  J. Peters,et al.  SCF and APC: the Yin and Yang of cell cycle regulated proteolysis. , 1998, Current opinion in cell biology.

[8]  A. Levine,et al.  Nuclear Export Is Required for Degradation of Endogenous p53 by MDM2 and Human Papillomavirus E6 , 1998, Molecular and Cellular Biology.

[9]  C. Brandl,et al.  TOM1p, a yeast hect-domain protein which mediates transcriptional regulation through the ADA/SAGA coactivator complexes. , 1998, Journal of molecular biology.

[10]  P. Leder,et al.  WW domain-mediated interactions reveal a spliceosome-associated protein that binds a third class of proline-rich motif: the proline glycine and methionine-rich motif. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[11]  M. Hochstrasser,et al.  Degradation Signal Masking by Heterodimerization of MATα2 and MATa1 Blocks Their Mutual Destruction by the Ubiquitin-Proteasome Pathway , 1998, Cell.

[12]  T. Südhof,et al.  C2-domains, Structure and Function of a Universal Ca2+-binding Domain* , 1998, The Journal of Biological Chemistry.

[13]  J. Corden,et al.  Ultraviolet Radiation-induced Ubiquitination and Proteasomal Degradation of the Large Subunit of RNA Polymerase II , 1998, The Journal of Biological Chemistry.

[14]  M. Sudol,et al.  The WW Domain of Neural Protein FE65 Interacts with Proline-rich Motifs in Mena, the Mammalian Homolog of DrosophilaEnabled* , 1997, The Journal of Biological Chemistry.

[15]  F. Portillo,et al.  Glucose activation of the yeast plasma membrane H+‐ATPase requires the ubiquitin–proteasome proteolytic pathway , 1997, FEBS letters.

[16]  J. P. Jensen,et al.  Subcellular Localization and Ubiquitin-conjugating Enzyme (E2) Interactions of Mammalian HECT Family Ubiquitin Protein Ligases* , 1997, The Journal of Biological Chemistry.

[17]  A. Sparks,et al.  Using Molecular Repertoires to Identify High-Affinity Peptide Ligands of the WW Domain of Human and Mouse YAP , 1997, Biological chemistry.

[18]  P. Leder,et al.  FBP WW domains and the Abl SH3 domain bind to a specific class of proline‐rich ligands , 1997, The EMBO journal.

[19]  J. Huibregtse,et al.  The large subunit of RNA polymerase II is a substrate of the Rsp5 ubiquitin-protein ligase. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[20]  R. Halaban,et al.  UV-induced ubiquitination of RNA polymerase II: a novel modification deficient in Cockayne syndrome cells. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[21]  M. Vigneron,et al.  In vivo degradation of RNA polymerase II largest subunit triggered by alpha-amanitin. , 1996, Nucleic acids research.

[22]  M. Imhof,et al.  Yeast RSP5 and its human homolog hRPF1 potentiate hormone-dependent activation of transcription by human progesterone and glucocorticoid receptors , 1996, Molecular and cellular biology.

[23]  B. André,et al.  Ubiquitination Mediated by the Npi1p/Rsp5p Ubiquitin-protein Ligase Is Required for Endocytosis of the Yeast Uracil Permease (*) , 1996, The Journal of Biological Chemistry.

[24]  F. R. Papa,et al.  The yeast SEN3 gene encodes a regulatory subunit of the 26S proteasome complex required for ubiquitin-dependent protein degradation in vivo , 1995, Molecular and cellular biology.

[25]  B. André,et al.  NPI1, an essential yeast gene involved in induced degradation of Gap1 and Fur4 permeases, encodes the Rsp5 ubiquitin—protein ligase , 1995, Molecular microbiology.

[26]  M. Sudol,et al.  The WW domain of Yes-associated protein binds a proline-rich ligand that differs from the consensus established for Src homology 3-binding modules. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[27]  M. Scheffner,et al.  A family of proteins structurally and functionally related to the E6-AP ubiquitin-protein ligase. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[28]  Martin Scheffner,et al.  Protein ubiquitination involving an E1–E2–E3 enzyme ubiquitin thioester cascade , 1995, Nature.

[29]  P. Hanawalt.  Transcription-coupled repair and human disease. , 1994, Science.

[30]  J. Hoeijmakers,et al.  Nucleotide excision repair. II: From yeast to mammals. , 1993, Trends in genetics : TIG.

[31]  P. Silver,et al.  Mutations that alter both localization and production of a yeast nuclear protein. , 1988, Genes & development.

[32]  P. Hanawalt,et al.  Selective removal of transcription-blocking DNA damage from the transcribed strand of the mammalian DHFR gene , 1987, Cell.

[33]  P. Howley,et al.  Ubiquitination of the p53 Tumor Suppressor , 1998 .

[34]  D. Bushnell,et al.  The Med proteins of yeast and their function through the RNA polymerase II carboxy-terminal domain. , 1998, Genes & development.

[35]  J. Peters,et al.  Ubiquitin and the Biology of the Cell , 1998, Springer US.

[36]  T. Südhof,et al.  C 2-domains , Structure and Function of a Universal Ca 2 1-binding Domain * , 1998 .

[37]  M. Hochstrasser.  Ubiquitin-dependent protein degradation. , 1996, Annual review of genetics.

[38]  P. Sung,et al.  DNA repair genes and proteins of Saccharomyces cerevisiae. , 1993, Annual review of genetics.

[39]  S. Jentsch.  The ubiquitin-conjugation system. , 1992, Annual review of genetics.