Novel telomere-anchored PCR approach for studying sexual stage telomeres in Aspergillus nidulans.

Telomere length varies between germline and somatic cells of the same organism, leading to the hypothesis that telomeres are lengthened during meiosis. However, little is known about the meiotic telomere length in many organisms. In the filamentous fungus Aspergillus nidulans, the telomere lengths i...

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Main Authors: Nengding Wang, Saajidha Rizvydeen, Mithaq Vahedi, Daysi M Vargas Gonzalez, Amanda L Allred, Dustin W Perry, Peter M Mirabito, Karen E Kirk
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2014-01-01
Series:PLoS ONE
Online Access:http://europepmc.org/articles/PMC4057176?pdf=render
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spelling doaj-64ec390a9db24c0b823f1d16bf1f1b502020-11-24T21:38:55ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0196e9949110.1371/journal.pone.0099491Novel telomere-anchored PCR approach for studying sexual stage telomeres in Aspergillus nidulans.Nengding WangSaajidha RizvydeenMithaq VahediDaysi M Vargas GonzalezAmanda L AllredDustin W PerryPeter M MirabitoKaren E KirkTelomere length varies between germline and somatic cells of the same organism, leading to the hypothesis that telomeres are lengthened during meiosis. However, little is known about the meiotic telomere length in many organisms. In the filamentous fungus Aspergillus nidulans, the telomere lengths in hyphae and asexual spores are invariant. No study using existing techniques has determined the telomere length of the sexual ascospores due to the relatively low abundance of pure meiotic cells in A. nidulans and the small quantity of DNA present. To address this, we developed a simple and sensitive PCR strategy to measure the telomere length of A. nidulans meiotic cells. This novel technique, termed "telomere-anchored PCR," measures the length of the telomere on chromosome II-L using a small fraction of the DNA required for the traditional terminal restriction fragment (TRF) Southern analysis. Using this approach, we determined that the A. nidulans ascospore telomere length is virtually identical to telomeres of other cell types from this organism, approximately 110 bp, indicating that a surprisingly strict telomere length regulation exists in the major cell types of A. nidulans. When the hyphal telomeres were measured in a telomerase reverse transcriptase (TERT) knockout strain, small decreases in length were readily detected. Thus, this technique can detect telomeres in relatively rare cell types and is particularly sensitive in measuring exceptionally short telomeres. This rapid and inexpensive telomere-anchored PCR method potentially can be utilized in other filamentous fungi and types of organisms.http://europepmc.org/articles/PMC4057176?pdf=render
collection DOAJ
language English
format Article
sources DOAJ
author Nengding Wang
Saajidha Rizvydeen
Mithaq Vahedi
Daysi M Vargas Gonzalez
Amanda L Allred
Dustin W Perry
Peter M Mirabito
Karen E Kirk
spellingShingle Nengding Wang
Saajidha Rizvydeen
Mithaq Vahedi
Daysi M Vargas Gonzalez
Amanda L Allred
Dustin W Perry
Peter M Mirabito
Karen E Kirk
Novel telomere-anchored PCR approach for studying sexual stage telomeres in Aspergillus nidulans.
PLoS ONE
author_facet Nengding Wang
Saajidha Rizvydeen
Mithaq Vahedi
Daysi M Vargas Gonzalez
Amanda L Allred
Dustin W Perry
Peter M Mirabito
Karen E Kirk
author_sort Nengding Wang
title Novel telomere-anchored PCR approach for studying sexual stage telomeres in Aspergillus nidulans.
title_short Novel telomere-anchored PCR approach for studying sexual stage telomeres in Aspergillus nidulans.
title_full Novel telomere-anchored PCR approach for studying sexual stage telomeres in Aspergillus nidulans.
title_fullStr Novel telomere-anchored PCR approach for studying sexual stage telomeres in Aspergillus nidulans.
title_full_unstemmed Novel telomere-anchored PCR approach for studying sexual stage telomeres in Aspergillus nidulans.
title_sort novel telomere-anchored pcr approach for studying sexual stage telomeres in aspergillus nidulans.
publisher Public Library of Science (PLoS)
series PLoS ONE
issn 1932-6203
publishDate 2014-01-01
description Telomere length varies between germline and somatic cells of the same organism, leading to the hypothesis that telomeres are lengthened during meiosis. However, little is known about the meiotic telomere length in many organisms. In the filamentous fungus Aspergillus nidulans, the telomere lengths in hyphae and asexual spores are invariant. No study using existing techniques has determined the telomere length of the sexual ascospores due to the relatively low abundance of pure meiotic cells in A. nidulans and the small quantity of DNA present. To address this, we developed a simple and sensitive PCR strategy to measure the telomere length of A. nidulans meiotic cells. This novel technique, termed "telomere-anchored PCR," measures the length of the telomere on chromosome II-L using a small fraction of the DNA required for the traditional terminal restriction fragment (TRF) Southern analysis. Using this approach, we determined that the A. nidulans ascospore telomere length is virtually identical to telomeres of other cell types from this organism, approximately 110 bp, indicating that a surprisingly strict telomere length regulation exists in the major cell types of A. nidulans. When the hyphal telomeres were measured in a telomerase reverse transcriptase (TERT) knockout strain, small decreases in length were readily detected. Thus, this technique can detect telomeres in relatively rare cell types and is particularly sensitive in measuring exceptionally short telomeres. This rapid and inexpensive telomere-anchored PCR method potentially can be utilized in other filamentous fungi and types of organisms.
url http://europepmc.org/articles/PMC4057176?pdf=render
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