Cargando…

Spatial and Temporal Dynamics of DNA Replication Sites in Mammalian Cells

Fluorescence microscopic analysis of newly replicated DNA has revealed discrete granular sites of replication (RS). The average size and number of replication sites from early to mid S-phase suggest that each RS contains numerous replicons clustered together. We are using fluorescence laser scanning...

Descripción completa

Detalles Bibliográficos
Autores principales: Ma, Hong, Samarabandu, Jagath, Devdhar, Rekandu S., Acharya, Raj, Cheng, Ping-chin, Meng, Chunling, Berezney, Ronald
Formato: Texto
Lenguaje:English
Publicado: The Rockefeller University Press 1998
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2132991/
https://www.ncbi.nlm.nih.gov/pubmed/9852140
_version_ 1782142569822552064
author Ma, Hong
Samarabandu, Jagath
Devdhar, Rekandu S.
Acharya, Raj
Cheng, Ping-chin
Meng, Chunling
Berezney, Ronald
author_facet Ma, Hong
Samarabandu, Jagath
Devdhar, Rekandu S.
Acharya, Raj
Cheng, Ping-chin
Meng, Chunling
Berezney, Ronald
author_sort Ma, Hong
collection PubMed
description Fluorescence microscopic analysis of newly replicated DNA has revealed discrete granular sites of replication (RS). The average size and number of replication sites from early to mid S-phase suggest that each RS contains numerous replicons clustered together. We are using fluorescence laser scanning confocal microscopy in conjunction with multidimensional image analysis to gain more precise information about RS and their spatial-temporal dynamics. Using a newly improved imaging segmentation program, we report an average of ∼1,100 RS after a 5-min pulse labeling of 3T3 mouse fibroblast cells in early S-phase. Pulse-chase-pulse double labeling experiments reveal that RS take ∼45 min to complete replication. Appropriate calculations suggest that each RS contains an average of 1 mbp of DNA or ∼6 average-sized replicons. Double pulse–double chase experiments demonstrate that the DNA sequences replicated at individual RS are precisely maintained temporally and spatially as the cell progresses through the cell cycle and into subsequent generations. By labeling replicated DNA at the G(1)/S borders for two consecutive cell generations, we show that the DNA synthesized at early S-phase is replicated at the same time and sites in the next round of replication.
format Text
id pubmed-2132991
institution National Center for Biotechnology Information
language English
publishDate 1998
publisher The Rockefeller University Press
record_format MEDLINE/PubMed
spelling pubmed-21329912008-05-01 Spatial and Temporal Dynamics of DNA Replication Sites in Mammalian Cells Ma, Hong Samarabandu, Jagath Devdhar, Rekandu S. Acharya, Raj Cheng, Ping-chin Meng, Chunling Berezney, Ronald J Cell Biol Article Fluorescence microscopic analysis of newly replicated DNA has revealed discrete granular sites of replication (RS). The average size and number of replication sites from early to mid S-phase suggest that each RS contains numerous replicons clustered together. We are using fluorescence laser scanning confocal microscopy in conjunction with multidimensional image analysis to gain more precise information about RS and their spatial-temporal dynamics. Using a newly improved imaging segmentation program, we report an average of ∼1,100 RS after a 5-min pulse labeling of 3T3 mouse fibroblast cells in early S-phase. Pulse-chase-pulse double labeling experiments reveal that RS take ∼45 min to complete replication. Appropriate calculations suggest that each RS contains an average of 1 mbp of DNA or ∼6 average-sized replicons. Double pulse–double chase experiments demonstrate that the DNA sequences replicated at individual RS are precisely maintained temporally and spatially as the cell progresses through the cell cycle and into subsequent generations. By labeling replicated DNA at the G(1)/S borders for two consecutive cell generations, we show that the DNA synthesized at early S-phase is replicated at the same time and sites in the next round of replication. The Rockefeller University Press 1998-12-14 /pmc/articles/PMC2132991/ /pubmed/9852140 Text en This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).
spellingShingle Article
Ma, Hong
Samarabandu, Jagath
Devdhar, Rekandu S.
Acharya, Raj
Cheng, Ping-chin
Meng, Chunling
Berezney, Ronald
Spatial and Temporal Dynamics of DNA Replication Sites in Mammalian Cells
title Spatial and Temporal Dynamics of DNA Replication Sites in Mammalian Cells
title_full Spatial and Temporal Dynamics of DNA Replication Sites in Mammalian Cells
title_fullStr Spatial and Temporal Dynamics of DNA Replication Sites in Mammalian Cells
title_full_unstemmed Spatial and Temporal Dynamics of DNA Replication Sites in Mammalian Cells
title_short Spatial and Temporal Dynamics of DNA Replication Sites in Mammalian Cells
title_sort spatial and temporal dynamics of dna replication sites in mammalian cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2132991/
https://www.ncbi.nlm.nih.gov/pubmed/9852140
work_keys_str_mv AT mahong spatialandtemporaldynamicsofdnareplicationsitesinmammaliancells
AT samarabandujagath spatialandtemporaldynamicsofdnareplicationsitesinmammaliancells
AT devdharrekandus spatialandtemporaldynamicsofdnareplicationsitesinmammaliancells
AT acharyaraj spatialandtemporaldynamicsofdnareplicationsitesinmammaliancells
AT chengpingchin spatialandtemporaldynamicsofdnareplicationsitesinmammaliancells
AT mengchunling spatialandtemporaldynamicsofdnareplicationsitesinmammaliancells
AT berezneyronald spatialandtemporaldynamicsofdnareplicationsitesinmammaliancells