Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2020 Nov;37(11):585-595.
doi: 10.1002/yea.3517. Epub 2020 Aug 15.

Simultaneous visualisation of the complete sets of telomeres from the MmeI generated terminal restriction fragments in yeasts

Jun Liu  1 , Xiaojing Hong  1 , Chao-Ya Liang  1 , Jun-Ping Liu  1   2   3
Affiliations

Simultaneous visualisation of the complete sets of telomeres from the MmeI generated terminal restriction fragments in yeasts

Jun Liu et al. Yeast. 2020 Nov.

Abstract

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, XhoI or PstI 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 MmeI. Based on analyses of the endonuclease cleavage sites, we found that the TRFs generated by MmeI displayed two distinguishable bands in the sizes of ~500 and ~700 bp comprising telomeres (300 bp) and subtelomeres (200-400 bp). The modified MmeI-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.

Keywords: Saccharomyces cerevisiae; Saccharomyces paradoxus; Southern blotting; X element; X-element-only telomere; Y′ element; Y′-element-containing telomere; restriction endonuclease MmeI; telomere; telomere length; terminal restriction fragment; yeast.

PubMed Disclaimer

Similar articles

See all similar articles

Cited by

References

REFERENCES

    1. Advani, V. M., Belew, A. T., & Dinman, J. D. (2013). Yeast telomere maintenance is globally controlled by programmed ribosomal frameshifting and the nonsense-mediated mRNA decay pathway. Translation (Austin), 1, e24418-e24418.
    1. Baird, D. M., Britt-Compton, B., Rowson, J., Amso, N. N., Gregory, L., & Kipling, D. (2005). Telomere instability in the male germline. Human Molecular Genetics, 15, 45-51.
    1. Berthiau, A. S., Yankulov, K., Bah, A., Revardel, E., Luciano, P., Wellinger, R. J., … Gilson, E. (2006). Subtelomeric proteins negatively regulate telomere elongation in budding yeast. The EMBO Journal, 25, 846-856. https://doi.org/10.1038/sj.emboj.7600975
    1. Boulton, S. J., & Jackson, S. P. (1996). Identification of a Saccharomyces cerevisiae Ku80 homologue: Roles in DNA double strand break rejoining and in telomeric maintenance. Nucleic Acids Research, 24, 4639-4648. https://doi.org/10.1093/nar/24.23.4639
    1. Boyd, A. C., Charles, I. G., Keyte, J. W., & Brammar, W. J. (1986). Isolation and computer-aided characterization of MmeI, a type II restriction endonuclease from Methylophilus methylotrophus. Nucleic Acids Research, 14, 5255-5274. https://doi.org/10.1093/nar/14.13.5255

Publication types

MeSH terms

Substances

LinkOut - more resources