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Mulder KW, et al.  (2007) Modulation of Ubc4p/Ubc5p-Mediated Stress Responses by the RING-Finger-Dependent Ubiquitin-Protein Ligase Not4p in Saccharomyces cerevisiae. Genetics 176(1):181-92

Abstract: The Ccr4-Not complex consists of nine subunits and acts as a regulator of mRNA biogenesis in Saccharomyces cerevisiae. The human ortholog of yeast NOT4, CNOT4, displays UbcH5B-dependent ubiquitin-protein ligase (E3 ligase) activity in a reconstituted in vitro system. However, an in vivo role for this enzymatic activity has not been identified. Site-directed mutagenesis of the RING finger of yeast Not4p identified residues required for interaction with Ubc4p and Ubc5p, the yeast orthologs of UbcH5B. Subsequent in vitro assays with purified Ccr4-Not complexes showed Not4p-mediated E3 ligase activity, which was dependent on the interaction with Ubc4p. To investigate the in vivo relevance of this activity, we performed synthetic genetic array (SGA) analyses using not4Delta and not4L35A alleles. This indicates involvement of the RING finger of Not4p in transcription, ubiquitylation, and DNA damage responses. In addition, we found a phenotypic overlap between deletions of UBC4 and mutants encoding single-amino-acid substitutions of the RING finger of Not4p. Together, our results show that Not4p functions as an E3 ligase by modulating Ubc4p/Ubc5p-mediated stress responses in vivo.

Status: Published Type: Journal Article PubMed ID: 17513889

Topics addressed in this paper

Number of different genes curated to this paper: 60

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Topics Genes linked to topics (#1 - 10 )
ARF1 BRO1 BUD19 BUL1 BUR2 CAF120 CAF40 CBF1 CCR4 DOA1
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Topics Genes linked to topics (#11 - 20 )
DOA4 ECM4 EST1 GAL11 GUF1 HSL7 HSP104 HSP42 HSP78 LOT5
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Topics Genes linked to topics (#21 - 30 )
MMS22 MOT2 MRT4 NNT1 NOT5 NPL4 OPI8 PGD1 PIB1 PPN1
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Topics Genes linked to topics (#31 - 40 )
PSH1 RAD6 RNR2 RNR3 RNR4 RPA49 SNF2 SOD1 SPT20 UBC4
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Topics Genes linked to topics (#41 - 50 )
UBC5 UBP6 URE2 VAM10 VMA11 VMA13 VMA4 VPS27 VPS38 VPS4
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Topics Genes linked to topics (#51 - 60 )
VPS5 VPS8 YDR417C YJL169W YKE2 YKR005C YLR232W YMR085W YOR302W YPL168W
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