Development of a two-part transcription probe to determine the completeness of temporal and spatial compartmentalization of gene expression during bacterial development

Proceedings of the National Academy of Sciences of the United States of America
Z Li, P J Piggot

Abstract

We have developed a two-part test, using the Bacillus subtilis sacB/SacY transcription antitermination system, to evaluate the completeness of temporal and spatial compartmentalization of gene expression during bacterial cell development. Transcription of sacY(1-55) (encoding a constitutively active form of the antiterminator, SacY) is directed by one promoter, whereas transcription of sacB'-'lacZ (the target of SacY action) is directed by the same or another promoter. To obtain beta-galactosidase activity, SacY(1-55) needs to be present when sacB'-'lacZ is being transcribed. We tested the system by analyzing the spatial compartmentalization of the activities of RNA polymerase final sigma factors, which are tightly regulated during sporulation of B. subtilis: final sigma(F) and then final sigma(G) in the prespore, final sigma(E) and then final sigma(K) in the mother cell. We have confirmed that the activities of final sigma(F) and final sigma(E) are spatially compartmentalized. We have demonstrated that there is also sharp temporal compartmentalization, with little or no overlap in the activities of final sigma(F) and final sigma(G) or of final sigma(E) and final sigma(K). In contrast, we found no compartmentalization of the ac...Continue Reading

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Citations

Mar 29, 2003·Journal of Molecular Biology·Patrick EichenbergerRichard Losick
Jan 28, 2003·Trends in Genetics : TIG·Jonathan Dworkin
Jul 3, 2008·Genes & Development·Kathleen A MarquisDavid Z Rudner
Dec 19, 2006·Journal of Bacteriology·Teresa CostaAdriano O Henriques
Jun 10, 2004·Microbiology and Molecular Biology Reviews : MMBR·David W Hilbert, Patrick J Piggot
Nov 24, 2004·Current Opinion in Microbiology·Patrick J Piggot, David W Hilbert
Sep 14, 2010·Current Biology : CB·Michiel J L de HoonDennis Vitkup
Dec 13, 2001·Developmental Cell·D Z Rudner, R Losick
Oct 30, 2016·Frontiers in Microbiology·Robyn T EijlanderOscar P Kuipers

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