Abstract
In the yeast two-hybrid system, the N-terminal region of Rap1p was shown to interact with Gcr1p and Gcr2p. Disruption of gcr1 and/or gcr2 in the two-hybrid reporter strain demonstrated that the interaction with Gcr1p does not require Gcr2p, whereas the interaction with Gcr2p is mediated through Gcr1p. Deletion of the N-terminal region of Rap1p alone did not show a growth phenotype, but a growth defect was observed when this mutation was combined with a gcr2 deletion. The poor growth of the gcr1 null mutant was not affected further by the N-terminal deletion of Rap1p, but the growth of gcr1 strains with mutations in the DNA binding region of Gcr1p was affected by the removal of the N-terminal region of Rap1p. These results suggest that one function of the N-terminal region of Rap1p, presumably the BRCT domain, is to facilitate the binding of Gcr1p to the promoter by a protein-protein interaction.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Blotting, Western
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DNA, Fungal / genetics
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DNA, Fungal / metabolism
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DNA-Binding Proteins / genetics
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DNA-Binding Proteins / metabolism*
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Fungal Proteins / genetics
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Fungal Proteins / metabolism*
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Gene Expression Regulation, Fungal / genetics
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Gene Expression Regulation, Fungal / physiology*
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Mutagenesis
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RNA, Fungal / chemistry
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RNA, Fungal / genetics
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Reverse Transcriptase Polymerase Chain Reaction
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Saccharomyces cerevisiae / genetics
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Saccharomyces cerevisiae / metabolism*
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Saccharomyces cerevisiae Proteins / genetics
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Saccharomyces cerevisiae Proteins / metabolism*
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Shelterin Complex
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Telomere-Binding Proteins / genetics
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Telomere-Binding Proteins / metabolism*
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Trans-Activators / genetics
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Trans-Activators / metabolism*
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Transcription Factors / genetics
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Transcription Factors / metabolism*
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Transcriptional Activation / genetics
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Transcriptional Activation / physiology*
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Two-Hybrid System Techniques
Substances
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DNA, Fungal
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DNA-Binding Proteins
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Fungal Proteins
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GCR1 protein, S cerevisiae
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GCR2 protein, S cerevisiae
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RAP1 protein, S cerevisiae
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RNA, Fungal
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Saccharomyces cerevisiae Proteins
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Shelterin Complex
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Telomere-Binding Proteins
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Trans-Activators
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Transcription Factors