Replication Termini in the RDNA of Synchronized Pea Root Cells (Pisum Sativum)

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 1988 Feb 1

PMID 16453826

Citations 12

Authors

P Hernandez

S S Lamm

C A Bjerknes

J V Hof

Affiliations

Soon will be listed here.

Abstract

In synchronized root cells of Pisum sativum (cv. Alaska) the joining of nascent replicons is delayed until cells reach the S-G(2) boundary or early G(2) phase. To determine if the delayed ligation of nascent chains occurs at specific termination sites, we mapped the location of arrested forks in the ribosomal DNA (rDNA) repeats from cells in late S and G(2) phases. Two-dimensional (neutral-alkaline) agarose electrophoresis and Southern blot hybridization with specific rDNA sequences show that only cells located at the S-G(2) boundary and early G(2) phase produce alkali-released rDNA fragments of discrete size. The released fragments are from a particular restriction fragment, demonstrating that the replication forks stop non-randomly within the rDNA repeats. Indirect end-labeling with probes homologous to one or the other end of the fork-containing restriction fragment shows that there are two termination regions, T(1) and T(2), where forks stop. T(1) is located in the non-transcribed spacer and T(2) is at the junction between the non-transcribed spacer and the 18S gene. The two termini are separated by 1.3 kb. Replication forks stop at identical sites in both the 8.6- and 9.0-kb rDNA repeat size classes indicating that these sites are sequence determined.

Citing Articles

Proximity of an ARS consensus sequence to a replication origin of pea (Pisum sativum).

Hernandez P, Bjerknes C, Lamm S, Vant Hof J Plant Mol Biol. 2013; 10(5):413-22.

PMID: 24277589 DOI: 10.1007/BF00014947.

Initiation of DNA replication: functional and evolutionary aspects.

Bryant J, Aves S Ann Bot. 2011; 107(7):1119-26.

PMID: 21508040 PMC: 3091809. DOI: 10.1093/aob/mcr075.

Interruption of Somatic Embryogenesis in Daucus carota L. by 5-Bromodeoxyuridine.

Thomas J, Nessler C, Katterman F Plant Physiol. 1989; 90(3):921-7.

PMID: 16666898 PMC: 1061821. DOI: 10.1104/pp.90.3.921.

Characterization of the pea rDNA replication fork barrier: putative cis-acting and trans-acting factors.

Schvartzman J, Krimer D, Hernandez P Plant Mol Biol. 1999; 40(1):99-110.

PMID: 10394949 DOI: 10.1023/a:1026405311132.

Expansion and contraction of ribosomal DNA repeats in Saccharomyces cerevisiae: requirement of replication fork blocking (Fob1) protein and the role of RNA polymerase I.

Kobayashi T, Heck D, Nomura M, Horiuchi T Genes Dev. 1998; 12(24):3821-30.

PMID: 9869636 PMC: 317266. DOI: 10.1101/gad.12.24.3821.

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