• Title/Summary/Keyword: Telomeric repeats

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Cytogenetic Mapping of Carthamus tinctorius L. with Tandemly Repeated DNA Sequences by Fluorescence in situ Hybridization

  • Mancia, Franklin Hinosa;Ju, Yoon Ha;Lim, Ki-Byung;Kim, Jung Sun;Nam, Sang Yong;Hwang, Yoon-Jung
    • Korean Journal of Plant Resources
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    • v.30 no.6
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    • pp.654-661
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    • 2017
  • Dual-color fluorescence in situ hybridization karyotype analysis was created using repetitive sequences including two types of rDNA repeats (45S and 5S rDNAs) and Arabidopsis-type telomere sequence repeats. The somatic metaphase cells of Carthamus tinctorius were observed as diploids (2n=2x=24). A symmetrical or slightly asymmetrical karyotype with seven pairs of metacentric and five pairs of submetacentric chromosomes was observed. The lengths of the somatic metaphase chromosomes ranged from 4.18 to $6.53{\mu}m$, with a total length of $60.71{\mu}m$. One locus of 45S rDNA was located on the pericentromeric regions of three pairs of chromosomes and the other pair was situated on the terminal regions of the short arms of a single pair of chromosomes. One locus of 5S rDNA was detected on the interstitial regions of the short arms of two pairs of chromosomes. Arabidopsis-type telomeric repeats were detected on the terminal regions of all pairs of chromosomes. Co-localization of loci between telomeric repeats and 45S rDNA was observed in a single pair of chromosomes. The results provide additional information for the existing physical mapping project of C. tinctorius and will also serve as a benchmark to a more intricate cytogenetic investigation of C. tinctorius and its related species.

Chromatin Structural Rearrangement during Dedifferentiation of Protoplasts of Cucumis sativus L.

  • Ondrej, Vladan;Kitner, Miloslav;Dolezalova, Ivana;Nadvornik, Petr;Navratilova, Bozena;Lebeda, Ales
    • Molecules and Cells
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    • v.27 no.4
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    • pp.443-447
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    • 2009
  • This paper reports on the structural rearrangement of satellite DNA type I repeats and heterochromatin during the dedifferentiation and cell cycling of mesophyll protoplasts of cucumber (Cucumis sativus). These repeats were localized in the telomeric heterochromatin of cucumber chromosomes and in the chromocenters of interphase nuclei. The dramatic reduction of heterochromatin involves decondensation of subtelomeric repeats in freshly isolated protoplasts; however, there are not a great many remarkable changes in the expression profile. In spite of that, reformation of the chromocenters, occurring 48 h after protoplast isolation, is accompanied by recondensation of satellite DNA type I; however, only partial reassembly of these repeats was revealed. In this study, FISH and a flow cytometry assay show a correlation between the partial chromocenter and the repeats reassembly, and with the reentry of cultivated protoplasts into the cell cycle and first cell division. After that, divided cells displayed a higher variability in the expression profile than did leaves' mesophyll cells and protoplasts.

Expression of Telomerase Activity and Apoptosis in Human Brain Tumors (인체 뇌종양조직에서 텔로머레이즈의 발현과 세포사멸)

  • Kim, Choong Hyun;Cheong, Jin Hwan;Bak, Koang Hum;Kim, Jae Min;Ko, Yong;Oh, Suck Jun
    • Journal of Korean Neurosurgical Society
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    • v.30 no.2
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    • pp.137-143
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    • 2001
  • Objective : Telomerase, a ribonucleoprotein adds telomere repeats to the ends of telomeres to compensate for the progressive loss. A favorable prognosis associated with low or no telomerase activity in some tumors, and cells transfected with antisense human telomerase lost telomeric repeats and die. We studied about the relationship between telomerase activity and apoptosis in the human brain tumors. Material and Methods : Between July 1998 and December 1999, 62 patients with brain tumors underwent surgery and their surgical specimens were obtained. Telomerase activity was investigated by telomeric repeats amplification protocol(TRAP) assay. Apoptosis was also evaluated by DNA fragmentation analysis. Differences and correlation in data were analyzed using Mann-Whitney test and Wilcoxon-signed rank test. Results : Expression rate of telomerase activity and apoptosis were 80% and 30% in malignant gliomas, 33% and 0% in low grade gliomas, 63% and 38% in meningiomas, 67% and 33% in pituitary adenomas, 33% and 33% in metastatic tumors, 67% and 17% in acoustic neurinomas, 100% and 100% in pineoblastomas, 100% and 0% in the hemangioblastoma, respectively. There was no significant difference of telomerase activity and apoptosis between histological types. But a significant difference was noted in the expression of telomerase activity between malignant gliomas and low grade gliomas(p = 0.022). Brain tumors with telomerase activity expressed the lower rate of apoptosis. A significant correlation was also found between telomerase activity and absence of apoptosis in the human brain tumors(p = 0.005). Conclusions : Our data suggests that telomerase may protect from apoptosis of the human brain tumors and also may play an important role in the biological malignancy of the gliomas.

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Age Prediction in the Chickens Using Telomere Quantity by Quantitative Fluorescence In situ Hybridization Technique

  • Kim, Y.J.;Subramani, V.K.;Sohn, S.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.5
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    • pp.603-609
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    • 2011
  • Telomeres are special structures at the ends of eukaryotic chromosomes. Vertebrate telomeres consist of tandem repeats of conserved TTAGGG sequence and associated proteins. Birds are interesting models for molecular studies on aging and cellular senescence because of their slow aging rates and longer life spans for their body size. In this longitudinal study, we explored the possibility of using telomeres as an age-marker to predict age in Single Comb White Leghorn layer chickens. We quantified the relative amount of telomeric DNA in isolated peripheral blood lymphocytes by the Quantitative Fluorescence in situ Hybridization technique on interphase nuclei (IQ FISH) using telomere-specific DNA probes. We found that the amount of telomeric DNA (ATD) reduced significantly with an increase in chronological age of the chicken. Especially, the telomere shortening rates are greatly increased in growing individuals compared to laying and old-aged individuals. Therefore, using the ATD values obtained by IQ FISH we established the possibility of age prediction in chickens based on the telomere theory of aging. By regression analysis of the ATD values at each age interval, we formulated an equation to predict the age of chickens. In conclusion, the telomeric DNA values by IQ FISH analyses can be used as an effective age-marker in predicting the chronological age of chickens. The study has implications in the breeding and population genetics of poultry, especially the reproductive potential.

Evaluation of Telomerase Inhibitors Using DE81 Filter Spotting Method from Natural Products

  • Lee, Sung-Jin;Woongchon Mar
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1998.11a
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    • pp.183-183
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    • 1998
  • Telomerase synthesizes telomeric DNA repeats onto chromosome ends de novo. Telomerase activation and telomere shortening in human somatic cells have been implicated in cell tumorigenesis and immortalization. In order to find the potential inhibitors against telomerase activitiy which can be used as potential anticancer agents, we screened about 100 kinds of natural products after partition into n-hexane, ethyl acetate and aqueous layers from methanol extracts. The inhibitory effects of these materials against telomerase enzyme activity were tested in 293T cell culture using telomeric repeat amplification protocol(TRAP). The incorporation of $\^$32/P-dGTP into amplified DNA was measured by adsorption to Whatman DE81 paper instead of using TRAP assay for screening the extracts of natural products. Strong effective compounds were not found in this study but DE81 filter spotting method may be a useful model for the screening. Some of the compounds which showed somewhat inhibitory effects had cytotoxic effects also.

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Cattle Age Prediction by Leukocytes Telomere Quantification (혈액세포의 텔로미어 함량을 이용한 소의 연령예측)

  • Choi, Na-Eun;Kim, Hyun-Sub;Choe, Chang-Yong;Jeon, Gwang-Joo;Sohn, Sea-Hwan
    • Journal of Animal Science and Technology
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    • v.52 no.5
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    • pp.367-374
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    • 2010
  • Telomeres at the end of chromosomes consist of tandem repeats of (TTAGGG)n DNA sequence and associated proteins. Telomeres have the essential functions in chromosome stability and genome integrity and are hence related to cell senescence and cancer. This study was carried out to quantify the amount of telomeric DNA and establish age prediction equations by using the quantity of telomeric DNA for cattle. Analysis of the telomere quantity of the lymphocytes was performed at different age, across breeds and between different sexes of cattle. We quantified the amount of telomeric DNA by the Q-FISH technique using the telomeric DNA probe in 460 cattle at age of 1~166 months in Korean Cattle and Holstein breeds. In results, we found that the amount of telomeric DNA decreased gradually with age. The amount of telomeric DNA of Korean Cattle was significantly higher than that of Holstein breed (P<0.01). In addition, the amount of telomeric DNA in male was significantly higher than that in female (P<0.01). Using the relationship between age and the amount of telomeric DNA in cattle, age predicting equations were established as a result of regression analysis. Because sex and breeds influenced telomeric DNA quantity, the age prediction equations were estimated separately in Korean Cattle females and Holstein females. The regression equations were $\hat{Y}$=$38.102X^2$-220.103X + 318.309 (P<0.0001, $R^2$=0.8019) in Korean Cattle females and $\hat{Y}$ = $42.799X^2$ - 199.682X + 242.106 (P<0.0001, $R^2$ = 0.8379) in Holstein females, where the X was quantity of telomeric DNA and Y was predicted age in months. These equations predicted the age of cattle with high significance and accuracy and have high R square values. Thus, it could be possible to scientifically predict the age using the above equations for Korean Cattle and Holstein females.

Shelterin Proteins and Cancer

  • Patel, Trupti NV;Vasan, Richa;Gupta, Divanshu;Patel, Jay;Trivedi, Manjari
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.8
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    • pp.3085-3090
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    • 2015
  • The telomeric end structures of the DNA are known to contain tandem repeats of TTAGGG sequence bound with specialised protein complex called the "shelterin complex". It comprises six proteins, namely TRF1, TRF2, TIN2, POT1, TPP1 and RAP1. All of these assemble together to form a complex with double strand and single strand DNA repeats at the telomere. Such an association contributes to telomere stability and its protection from undesirable DNA damage control-specific responses. However, any alteration in the structure and function of any of these proteins may lead to undesirable DNA damage responses and thus cellular senescence and death. In our review, we throw light on how mutations in the proteins belonging to the shelterin complex may lead to various malfunctions and ultimately have a role in tumorigenesis and cancer progression.

Amount of Telomeric DNA on Lymphocytes in Senescence Mouse by Quantitative Fluorescence in situ Hybridization (노화촉진마우스의 텔로미어 함량 분석)

  • Lee, Mi-Rang;Do, Kyoung-Tag;Han, Jyung-Ju;Moon, So-Hyun;Kang, Han-Seok;Kim, Seon-Ku;Shin, Teak-Soon;Lee, Hong-Goo;Hwang, Dae-Yon;Kim, Yong-Gyun;Sohn, Sea-Hwan;Choi, Na-Eun;Kim, Byeong-Woo;Cho, Byung-Wook
    • Journal of Life Science
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    • v.19 no.10
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    • pp.1463-1467
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    • 2009
  • Telomeres, comprised of tandem repeats of TTAGGG sequences, are special nucleoprotein structures that protect and stabilize chromosome ends. These structures form the crux of the telomere concept of aging, senescence and genomic instability. The classic terminal restriction fragment (TRF) analysis to quantify the amount of telomeric DNA is disadvantageous in species containing ultra long telomeres like in mice (100Kb). In this study, we used a more sensitive quantitative fluorescence in situ hybridization (Q FISH) technique to quantify telomeric DNA, and used it as a biological aging marker in mice. 12 litters each of Senescence-Resistant (SAMR1) and -Prone (SAMP1) known as senescence accelerated mouse strains were purchased from Central Lab, Animal Inc. We quantified the amount of telomeric DNA using telomere specific DNA probes on the two strains of male mice at 8 weeks, 18 weeks and 26 weeks of age. The amount of telomeric DNA correlated with aging and age associated changes in body and organ weight between SAMR1 and SAMP1 strains of mice. These data suggest the usefulness of the amount of telomeric DNA as a biological aging marker in human aging studies.

Telomerase reverse transcriptase in the regulation of gene expression

  • Zhou, Junzhi;Ding, Deqiang;Wang, Miao;Cong, Yu-Sheng
    • BMB Reports
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    • v.47 no.1
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    • pp.8-14
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    • 2014
  • Telomerase plays a pivotal role in the pathology of aging and cancer by maintaining genome integrity, controlling cell proliferation, and regulating tissue homeostasis. Telomerase is essentially composed of an RNA component, Telomerase RNA or TERC, which serves as a template for telomeric DNA synthesis, and a catalytic subunit, telomerase reverse transcriptase (TERT). The canonical function of TERT is the synthesis of telomeric DNA repeats, and the maintenance of telomere length. However, accumulating evidence indicates that TERT may also have some fundamental functions that are independent of its enzymatic activity. Among these telomere-independent activities of hTERT, the role of hTERT in gene transcription has been investigated in detail. Transcriptional regulation is a fundamental process in biological systems. Several studies have shown a direct involvement of hTERT in gene transcription. This mini-review will focus on the role of hTERT in gene transcription regulation, and discuss its possible mechanisms.