Chromosomal replication initiates and terminates at random sequences but at regular intervals in the ribosomal DNA of Xenopus early embryos.
We have analysed the replication of the chromosomal ribosomal DNA (rDNA) cluster in Xenopus embryos before the midblastula transition. Two-dimensional gel analysis showed that replication forks are associated with the nuclear matrix, as in differentiated cells, and gave no evidence for single-stranded replication intermediates (RIs). Bubbles, simple forks and double Ys were found in each restriction fragment analysed, showing that replication initiates and terminates without detectable sequence specificity. Quantification of the results and mathematical analysis showed that the average rDNA replicon replicates in 7.5 min and is 9-12 kbp in length. This time is close to the total S phase duration, and this replicon size is close to the maximum length of DNA which can be replicated from a single origin within this short S phase. We therefore infer that (i) most rDNA origins must be synchronously activated soon in S phase and (ii) origins must be evenly spaced, in order that no stretch of chromosomal DNA is left unreplicated at the end of S phase. Since origins are not specific sequences, it is suggested that this spatially and temporally concerted pattern of initiation matches some periodic chromatin folding, which itself need not rely on DNA sequence.
Hyrien
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Unité d'Embryologie Moléculaire, Institut Jacques Monod, Paris, France.
Méchali
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M
D000818:Animals; D001755:Blastocyst / Q000378:metabolism; D002875:Chromosomes / Q000378:metabolism; D004261:DNA Replication; D004275:DNA, Ribosomal / Q000096:biosynthesis* / Q000235:genetics; D015180:Electrophoresis, Gel, Two-Dimensional; D010957:Plasmids; D015183:Restriction Mapping; D014982:Xenopus laevis
D004275:DNA, Ribosomal