SUMMARY: Six mutants of Bacillus subtilis 168 that are temperature-sensitive in spore outgrowth were isolated. The outgrowth process proceeds normally at 35 °C, but at the non-permissive temperature (47 °C) it is arrested at a specific stage characteristic for each mutant strain. The mutants are not altered in vegetative growth whether at 35 °C or at 47 °C. They were characterized for their ability to synthesize RNA, proteins and DNA during outgrowth. A mutant defective in spore germination was also isolated; less than 5% of its spores can germinate at any of the temperatures tested. The mutations were mapped by means of transduction and transformation. The isolation of a number of outgrowth mutants which map at different loci and which affect outgrowth at different times is discussed in relation to the regulation of this process.
AdelbergE. A.,
MandelM.,
ChenG. C. C.1965; Optimal conditions for mutagenesis by N-methyl-Nʹ-nitro-N-nitrosoguanidine in Escherichia coli K12. Biochemical and Biophysical Research Communications 18:788–795
AlbertiniA. M.,
GalizziA.1975; Mutant of Bacillus subtilis with a temperature-sensitive lesion in ribonucleic acid synthesis during germination. Journal of Bacteriology 124:14–25
ArmstrongR. L.,
SueokaN.1968; Phase transition in ribonucleic acid synthesis during germination of B. subtilis spores. Proceedings of the National Academy of Sciences of the United States of America 59:153–160
BalassaG.1969; Biochemical genetics of bacterial sporulation. I. Unidirectional pleiotropic interactions among genes controlling sporulation in Bacillus subtilis. Molecular and General Genetics 104:73–103
CiarrocchiG.,
AttoliniC.,
CobianchiF.,
RivaS.,
FalaschiA.1977; Modulation of deoxyribonucleic acid polymerase III level during the life cycle of Bacillus subtilis. Journal of Bacteriology 131:776–783
CooteJ. G.,
MandelstamJ.1973; Use of constructed double mutants for determining the temporal order of expression of sporulation genes in Bacillus subtilis. Journal of Bacteriology 114:1254–1263
DawesI. W.,
HalvorsonH. O.1974; Temperature-sensitive mutants of Bacillus subtilis defective in spore outgrowth. Molecular and General Genetics 131:147–157
GalizziA.,
GorriniG.,
RollierA.,
PolsinelliM.1973; Mutants of Bacillus subtilis temperature sensitive in the outgrowth phase of spore germination. Journal of Bacteriology 113:1482–1490
GalizziA., . SiccardiA. G.,
AlbertiniA. M.,
AmileniA. R.,
MeneguzziG.,
PolsinelliM.1975; Properties of Bacillus subtilis mutants temperature sensitive in germination. Journal of Bacteriology 121:450–454
HochJ. A.,
BaratM.,
AnagnostopoulosC.1967; Transformation and transduction in recombination defective mutants of Bacillus subtilis. Journal of Bacteriology 93:1925–1937
KennettR. N.,
SueokaN.1971; Gene expression during outgrowth of B. subtilis spores. The relationship between order on the chromosome and temporal sequence of enzyme synthesis. Journal of Molecular Biology 60:31–44
KobayashiY.,
SteinbergW.,
HigaA.,
HalvorsonH. O.,
LevinthalC.1965; Sequential synthesis of macromolecules during outgrowth of bacterial spores.. In Spores III, pp. 200–212 Edited by
CampbellL. L.,
HalvorsonH. O.
Washington D. C.: American Society for Microbiology;
Lepesant-KejzlarovaJ.,
LepesantJ. A.,
WalleJ.,
BillaultA.,
DedonderR.1975; Revision of the linkage map of Bacillus subtilis 168: indications for circularity of the chromosome. Journal of Bacteriology 121:823–834
MandelstamJ.1976; Bacterial sporulation: a problem in the biochemistry and genetics of a primitive developmental system. Proceedings of the Royal Society of London B193:89–106
MazzaG.,
GalizziA.,
MinghettiA.,
SiccardiA. G.1973; Interaction between deoxyribonucleic acid and distamycin A studied by transformation in Bacillus subtilis. Antimicrobial Agents and Chemotherapy 3:384–391
NukushinaJ.,
IkedaY.1969; Genetic analysis of the developmental processes during germination and outgrowth of B. subtilis spores with temperature-sensitive mutants. Genetics 63:63–74
SiccardiA. G.,
GalizziA.,
MazzaG.,
ClivioA.,
AlbertiniA. M.1975a; Synchronous germination and outgrowth of fractionated B. subtilis spores: tool for the analysis of differentiation and division of bacterial cells. Journal of Bacteriology 121:13–19
SiccardiA. G.,
LanzaE.,
NielsenE.,
GalizziA.,
MazzaG.1975b; Genetic and physiological studies on the site of action of Distamycin A. Antimicrobial Agents and Chemotherapy 8:370–376
SpizizenJ.1958; Transformation of biochemically deficient strains of Bacillus subtilis by deoxy- ribonucleate. Proceedings of the National Academy of Sciences of the United States of America 44:1072–1078
SteinbergW.,
HalvorsonH. O.1968a; Timing of enzyme synthesis during outgrowth of Bacillus cereus. I. Ordered enzyme synthesis. Journal of Bacteriology 95:469–478
SteinbergW.,
HalvorsonH. O.1968b; Timing of enzyme synthesis during outgrowth of Bacillus cereus. II. Relationship between ordered enzyme synthesis and deoxyribonucleic acid replication. Journal of Bacteriology 95:479–489
TrowsdaleJ.,
SmithD. A.,
GouldG. W.1974; Mapping of germination mutants of Bacillus subtilis 168.. In Spore Research 1973, pp. 103–117 Edited by
BarkerA. N.,
GouldG. W.,
WolfJ.
London: Academic Press;
WaitesW. M.,
KayD.,
DawesI. W.,
WoodD. A.,
WarrenS. C.,
MandelstamJ.1970; Sporulation of Bacillus subtilis: correlation of biochemical events with morphological changes in asporogenous mutants. Biochemical Journal 118:667–676
WoeseC. R.,
VaryJ. C.,
HalvorsonH. O.1968; A kinetic model for bacterial spore germination. Proceedings of the National Academy of Sciences of the United States of America 59:869–875
YoshikawaH.1965; DNA synthesis during germination of Bacillus subtilis spores. Proceedings of the National Academy of Sciences of the United States of America 53:1476–1482
YoungF. E.,
WilsonG. A.1972; Genetics of Bacillus subtilis and other gram-positive sporu- lating bacilli.. In Spores V, pp. 77–106 Edited by
HalvorsonH. O.,
HansonR.,
CampbellL. L.
Washington, D. C.: American Society for Microbiology;