An essential yeast gene encoding a TTAGGG repeat-binding protein

C. Brigati, S. Kurtz, D. Balderes, G. Vidali, D. Shore

Research output: Contribution to journalArticlepeer-review

64 Scopus citations

Abstract

A yeast gene encoding a DNA-binding protein that recognizes the telomeric repeat sequence TTAGGG found in multicellular eukaryotes was identified by screening a λgt11 expression library with a radiolabeled TTAGGG multimer. This gene, which we refer to as TBF1 (TTAGGG repeat-binding factor 1), encodes a polypeptide with a predicted molecular mass of 63 kDa. The TBF1 protein, produced in vitro by transcription and translation of the cloned gene, binds to (TTAGGG)(n) probes and to a yeast telomeric junction sequence that contains two copies of the sequence TTAGGG separated by 5 bp. TBF1 appears to be identical to a previously described yeast TTAGGG-repeat binding activity called TBFα. TBF1 produced in vitro yields protein-DNA complexes with (TTAGGG)(n) probes that have mobilities on native polyacrylamide gels identical to those produced by partially purified TBFα from yeast cells. Furthermore, when extracts are prepared from a strain containing a TBF1 gene with an antigen tag, we find that the antigen copurifies with the predominant (TTAGGG)(n)-binding activity in the extracts. The DNA sequence of TBF1 was determined. The predicted protein sequence suggests that TBF1 may contain a nucleotide-binding domain, but no significant similarities to any other known proteins were identified, nor was an obvious DNA-binding motif apparent. Diploid cells heterozygous for a tbf1::URA3 insertion mutation are viable but upon sporulation give rise to tetrads with only two viable spores, both of which are Ura-, indicating that the TBF1 gene is essential for growth. Possible functions of TBF1 (TFBα) are discussed in light of these new results.

Original languageEnglish (US)
Pages (from-to)1306-1314
Number of pages9
JournalMolecular and cellular biology
Volume13
Issue number2
DOIs
StatePublished - 1993

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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