Breakdown and quantitation of the forked termination of replication intermediate of Bacillus subtilis

P. J B Hanley*, C. M. Carrigan, D. B. Rowe, R. G. Wake

*Corresponding author for this work

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Using a procedure that minimizes shear forces, the BamHI-derived forked termination of replication intermediate of Bacillus subtilis, called band I DNA, can be extracted with little or no accompanying band II DNA. It has been shown that band II DNA is a product of band I breakdown. Nuclease P1-mediated breakdown of the forked band I DNA proceeds in two steps. The first causes the release of one of the arms as band II DNA; in the second step, the remaining arm is cleaved away to yield the free stem. It is concluded that band I represents the primary termination of replication intermediate. A quantitative assessment of the level of band I in DNA from cells of the merodiploid strain, GSY1127, growing at different rates has been made. For cells grown in a minimal medium, at least, the experimentally measured level of band I is of the order (approx. 60%) of that predicted for a complete block to movement of the clockwise fork at the replication terminus, erC.

Original languageEnglish
Pages (from-to)721-727
Number of pages7
JournalJournal of molecular biology
Volume196
Issue number3
DOIs
Publication statusPublished - 5 Aug 1987
Externally publishedYes

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