Nucleic Acids Research, 2000, Vol. 28, No. 23 4733-4741
© 2000 Oxford University Press
Telomere-binding and Stn1p-interacting activities are required for the essential function of Saccharomyces cerevisiae Cdc13p
Institute of Biopharmaceutical Science, National Yang-Ming University, Shih-Pai, 112, Taipei, Taiwan, Republic of China
Yeast Saccharomyces cerevisiae Cdc13p is the telomere-binding protein that protects telomeres and regulates telomere length. It is documented that Cdc13p binds specifically to single-stranded TG1–3 telomeric DNA sequences and interacts with Stn1p. To localize the region for single-stranded TG1–3 DNA binding, Cdc13p mutants were constructed by deletion mutagenesis and assayed for their binding activity. Based on in vitro electrophoretic mobility shift assay, a 243-amino-acid fragment of Cdc13p (amino acids 451–693) was sufficient to bind single-stranded TG1–3 with specificity similar to that of the native protein. Consistent with the in vitro observation, in vivo one-hybrid analysis also indicated that this region of Cdc13p was sufficient to localize itself to telomeres. However, the telomere-binding region of Cdc13p (amino acids 451–693) was not capable of complementing the growth defects of cdc13 mutants. Instead, a region comprising the Stn1p-interacting and telomere-binding region of Cdc13p (amino acids 252–924) complemented the growth defects of cdc13 mutants. These results suggest that binding to telomeres by Cdc13p is not sufficient to account for the cell viability, interaction with Stn1p is also required. Taken together, we have defined the telomere-binding domain of Cdc13p and showed that both binding to telomeres and Stn1p by Cdc13p are required to maintain cell growth.
* To whom correspondence should be addressed. Tel: +86 2 2826 7258; Fax: +86 2 2820 0067; Email: jjlin{at}ym.edu.tw
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  C.-L. Hsu, Y.-S. Chen, S.-Y. Tsai, P.-J. Tu, M.-J. Wang, and J.-J. Lin Interaction of Saccharomyces Cdc13p with Pol1p, Imp4p, Sir4p and Zds2p is involved in telomere replication, telomere maintenance and cell growth control Nucleic Acids Res., January 23, 2004; 32(2): 511 - 521. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Enomoto, L. Glowczewski, J. Lew-Smith, and J. G. Berman Telomere Cap Components Influence the Rate of Senescence in Telomerase-Deficient Yeast Cells Mol. Cell. Biol., January 15, 2004; 24(2): 837 - 845. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. M. Anderson, W. A. Halsey, and D. S. Wuttke Delineation of the high-affinity single-stranded telomeric DNA-binding domain of Saccharomyces cerevisiae Cdc13 Nucleic Acids Res., October 1, 2002; 30(19): 4305 - 4313. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. L. DuBois, Z. W. Haimberger, M. W. McIntosh, and D. E. Gottschling A Quantitative Assay for Telomere Protection in Saccharomyces cerevisiae Genetics, July 1, 2002; 161(3): 995 - 1013. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J.-L. Mergny, J.-F. Riou, P. Mailliet, M.-P. Teulade-Fichou, and E. Gilson Natural and pharmacological regulation of telomerase Nucleic Acids Res., February 15, 2002; 30(4): 839 - 865. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y.-C. Lin, J.-W. Shih, C.-L. Hsu, and J.-J. Lin Binding and Partial Denaturing of G-quartet DNA by Cdc13p of Saccharomyces cerevisiae J. Biol. Chem., December 7, 2001; 276(50): 47671 - 47674. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Menez, E. Remy, and R. H. Buckingham Suppression of thermosensitive peptidyl-tRNA hydrolase mutation in Escherichia coli by gene duplication Microbiology, June 1, 2001; 147(6): 1581 - 1589. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y.-C. Lin, C.-L. Hsu, J.-W. Shih, and J.-J. Lin Specific Binding of Single-stranded Telomeric DNA by Cdc13p of Saccharomyces cerevisiae J. Biol. Chem., June 29, 2001; 276(27): 24588 - 24593. [Abstract] [Full Text] [PDF]
|
|