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Simultaneous visualisation of the complete sets of telomeres from the Mme I generated terminal restriction fragments in yeasts
Author(s) -
Liu Jun,
Hong Xiaojing,
Liang ChaoYa,
Liu JunPing
Publication year - 2020
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.3517
Subject(s) - subtelomere , telomere , biology , saccharomyces cerevisiae , restriction enzyme , restriction fragment , genetics , endonuclease , southern blot , minichromosome , yeast , dna , genome , microbiology and biotechnology , gene
Telomere length is measured using Southern blotting of the chromosomal terminal restriction fragments (TRFs) released by endonuclease digestion in cells from yeast to human. In the budding yeast Saccharomyces cerevisiae , Xho I or Pst I is applied to cut the subtelomere Y′ element and release TRFs from the 17 subtelomeres. However, telomeres from other 15 X‐element‐only subtelomeres are omitted from analysis. Here, we report a method for measuring all 32 telomeres in S. cerevisiae using the endonuclease Mme I. Based on analyses of the endonuclease cleavage sites, we found that the TRFs generated by Mme I displayed two distinguishable bands in the sizes of ~500 and ~700 bp comprising telomeres (300 bp) and subtelomeres (200–400 bp). The modified Mme I‐restricted TRF (mTRF) method recapitulated telomere shortening and lengthening caused by deficiencies of YKu and Rif1 respectively in S. cerevisiae . Furthermore, we found that mTRF was also applicable to telomere length analysis in S. paradoxus strains. These results demonstrate a useful tool for simultaneous detection of telomeres from all chromosomal ends with both X‐element‐only and Y′‐element subtelomeres in S. cerevisiae species.