Thermosensitive Bacillus Subtilis Mutants Which Lyse at the Non-permissive Temperature

Abstract

A collection of 655 thermosensitive mutants of Bacillus subtilis 168, obtained by indirect selection, was screened for those lysing at the non-permissive temperature. Thirty-three mutations thus identified were distributed by transformation into eight linkage groups designated lssA to lssH. The distribution was non-random. With the exception of group A, all groups were small, suggesting that mutations identified in each of them may map in one gene only. Linkage groups identified here were mapped in four different regions of the B. subtilis chromosome and their positions relative to reference markers were the following: (i) aroI-lssA-dal-purB; (ii) metC-lssB-lssC-furA-pyrE-cysC-lssD; (iii) lssF-gtaA-lssG-hisA-lssH-cysB; and (iv) cysA-lssE-dnaC-purA. Kinetics of N-acetyl-D-[1-14C]glucosamine incorporation revealed that groups A, B, C, D and F are deficient in peptidoglycan synthesis at the restrictive temperature. In group G, anomalies at the cell wall level were suggested by incorporation and growth curves. It appears that in almost all known cases, thermosensitive lysis mutations in B. subtilis either affect genes involved in peptidoglycan synthesis or lead, more or less directly, to induction of prophages.

The N -Acetylmannosamine Transferase Catalyzes the First Committed Step of Teichoic Acid Assembly in Bacillus subtilis and Staphylococcus aureus

“…Work on the essential nature of wall teichoic acid dates back many years to the discovery and characterization of temperature-sensitive B. subtilis tag mutants for poly(glycerol phosphate) synthesis by D. Karamata's lab (4,5,19). That work and follow-up studies by our research group (2,3,20) showed convincingly that genetic lesions in several wall teichoic acid biosynthetic steps led to cell death in vitro.…”

“…Wall teichoic acid has long been held as an essential component of the cell wall architecture (2)(3)(4)(5)19). However, recently, our group has demonstrated a complex pattern of dispensability for wall teichoic acid biosynthetic genes of both Bacillus subtilis and Staphylococcus aureus (9,10).…”

Incorporation of [2-3H]glycerol into cell surface components of Bacillus subtilis 168 and thermosensitive mutants affected in wall teichoic acid synthesis: effect of tunicamycin

“…exclusively concerned in the synthesis of PG precursors, do not develop swollen cells. In B. subtilis (Brandt & Karamata, 1987) as well as in E. coli (Mengin-Lecreulx & van Heijenoort, 1996), they are all associated with cell lysis.…”

Bacillus subtilis possesses a second determinant with extensive sequence similarity to the Escherichia coli mreB morphogene

“…Bacterial strains and plasmids utilized in this study are presented in Table 1. Plasmid pBK32, which contains a 4-kb Sau3A-to-SacI B. subtilis DNA fragment bearing spoIIID and mbl, was generously provided by R. Losick (Harvard University) (4,18). To obtain chromosomal DNA extending beyond the SacI site, the 1.2-kb PstI-to-SacI fragment bearing the 3Ј end of mbl ( Fig.…”