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Analysis of the RAP1 protein binding to homogeneous telomeric repeats in Saccharomyces castellii.

Wahlin, Johan LU and Cohn, Marita LU (2002) In Yeast 19(3). p.241-256
Abstract
The repressor activator protein 1 (RAP1) plays a role in telomere structure and function inS. cerevisiae. Here, the RAP1 homologue was identified and cloned from the budding yeast Saccharomyces castellii (scasRAP1). The scasRAP1 gene encodes a protein of 826 amino acids and shares an overall high degree of similarity with the S. cerevisiae RAP1 (scerRAP1). We demonstrate that the scasRAP1 is able to complement scerRAP1 in temperature-sensitive S. cerevisiae strains and is able to function as a regulator to maintain the original telomere lengths. Binding analyses of the E. coli-expressed scasRAP1 protein demonstrate that it needs two consecutive telomeric repeats in order to bind the S. castellii telomeric DNA sequences, and that it binds... (More)
The repressor activator protein 1 (RAP1) plays a role in telomere structure and function inS. cerevisiae. Here, the RAP1 homologue was identified and cloned from the budding yeast Saccharomyces castellii (scasRAP1). The scasRAP1 gene encodes a protein of 826 amino acids and shares an overall high degree of similarity with the S. cerevisiae RAP1 (scerRAP1). We demonstrate that the scasRAP1 is able to complement scerRAP1 in temperature-sensitive S. cerevisiae strains and is able to function as a regulator to maintain the original telomere lengths. Binding analyses of the E. coli-expressed scasRAP1 protein demonstrate that it needs two consecutive telomeric repeats in order to bind the S. castellii telomeric DNA sequences, and that it binds adjacent sites having a 16 bp centre-to-centre spacing. The binding affinity to telomeric DNA of several other yeasts is similar to that of scerRap1p. However, in contrast to scerRap1p, scasRap1p was found to bind the human telomeric sequence. Moreover, the scasRap1p was found to incorporate a variant repeat in its binding to the otherwise homogeneous telomeric DNA of S. castellii. This ability to bind various sites differing in DNA sequence indicates a high degree of adjustability in the binding of scasRap1p to DNA. (Less)
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organization
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Contribution to journal
publication status
published
subject
keywords
Telomere/*metabolism, Support, Amino Acid, Sequence Homology, Saccharomyces/*genetics/metabolism, Repressor Proteins/chemistry/genetics/*metabolism, Protein Binding, Polymerase Chain Reaction, Molecular Sequence Data, Electrophoretic Mobility Shift Assay, Deoxyribonuclease I/chemistry, Genetic Complementation Test, Non-U.S. Gov't, DNA-Binding Proteins/chemistry/genetics/*metabolism, DNA Footprinting, Molecular, Cloning, Amino Acid Sequence
in
Yeast
volume
19
issue
3
pages
241 - 256
publisher
John Wiley & Sons
external identifiers
  • wos:000174109200006
  • pmid:11816032
  • scopus:0036187742
ISSN
1097-0061
DOI
10.1002/yea.816
language
English
LU publication?
yes
id
e861aeb5-c038-4828-8d5d-fd0574617a05 (old id 106341)
alternative location
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11816032&dopt=Abstract
date added to LUP
2007-07-23 13:33:34
date last changed
2017-09-03 04:36:30
@article{e861aeb5-c038-4828-8d5d-fd0574617a05,
  abstract     = {The repressor activator protein 1 (RAP1) plays a role in telomere structure and function inS. cerevisiae. Here, the RAP1 homologue was identified and cloned from the budding yeast Saccharomyces castellii (scasRAP1). The scasRAP1 gene encodes a protein of 826 amino acids and shares an overall high degree of similarity with the S. cerevisiae RAP1 (scerRAP1). We demonstrate that the scasRAP1 is able to complement scerRAP1 in temperature-sensitive S. cerevisiae strains and is able to function as a regulator to maintain the original telomere lengths. Binding analyses of the E. coli-expressed scasRAP1 protein demonstrate that it needs two consecutive telomeric repeats in order to bind the S. castellii telomeric DNA sequences, and that it binds adjacent sites having a 16 bp centre-to-centre spacing. The binding affinity to telomeric DNA of several other yeasts is similar to that of scerRap1p. However, in contrast to scerRap1p, scasRap1p was found to bind the human telomeric sequence. Moreover, the scasRap1p was found to incorporate a variant repeat in its binding to the otherwise homogeneous telomeric DNA of S. castellii. This ability to bind various sites differing in DNA sequence indicates a high degree of adjustability in the binding of scasRap1p to DNA.},
  author       = {Wahlin, Johan and Cohn, Marita},
  issn         = {1097-0061},
  keyword      = {Telomere/*metabolism,Support,Amino Acid,Sequence Homology,Saccharomyces/*genetics/metabolism,Repressor Proteins/chemistry/genetics/*metabolism,Protein Binding,Polymerase Chain Reaction,Molecular Sequence Data,Electrophoretic Mobility Shift Assay,Deoxyribonuclease I/chemistry,Genetic Complementation Test,Non-U.S. Gov't,DNA-Binding Proteins/chemistry/genetics/*metabolism,DNA Footprinting,Molecular,Cloning,Amino Acid Sequence},
  language     = {eng},
  number       = {3},
  pages        = {241--256},
  publisher    = {John Wiley & Sons},
  series       = {Yeast},
  title        = {Analysis of the RAP1 protein binding to homogeneous telomeric repeats in Saccharomyces castellii.},
  url          = {http://dx.doi.org/10.1002/yea.816},
  volume       = {19},
  year         = {2002},
}