Temperature Sensitivity of Cell Growth in Escherichia coli associated with the Temperature Sensitive R(KM) Factor

Nature ◽  
1968 ◽  
Vol 219 (5151) ◽  
pp. 284-285 ◽  
Author(s):  
YOSHIRO TERAWAKI ◽  
YOSHIHIRO KAKIZAWA ◽  
HISAO TAKAYASU ◽  
MASANOSUKE YOSHIKAWA
Keyword(s):  
Escherichia Coli ◽  
Cell Growth ◽  
Temperature Sensitivity ◽  
Temperature Sensitive

Temperature-sensitive mutants of MscL mechanosensitive channel

2019 ◽  
Vol 166 (3) ◽  
pp. 281-288 ◽  
Author(s):  
Naoto Owada ◽  
Megumi Yoshida ◽  
Kohei Morita ◽  
Kenjiro Yoshimura
Keyword(s):  
Escherichia Coli ◽  
Amino Acids ◽  
Structural Change ◽  
Cell Growth ◽  
Thermodynamic Stability ◽  
Transmembrane Helix ◽  
Mechanosensitive Channel ◽  
Temperature Sensitive ◽  
Temperature Sensitive Mutants ◽  
Membrane Stretching

Abstract MscL is a mechanosensitive channel that undergoes a global conformational change upon application of membrane stretching. To elucidate how the structural stability and flexibility occur, we isolated temperature-sensitive (Ts) mutants of Escherichia coli MscL that allowed cell growth at 32°C but not at 42°C. Two Ts mutants, L86P and D127V, were identified. The L86P mutation occurred in the second transmembrane helix, TM2. Substitution of residues neighbouring L86 with proline also led to a Ts mutation, but the substitution of L86 with other amino acids did not result in a Ts phenotype, indicating that the Ts phenotype was due to a structural change of TM2 helix by the introduction of a proline residue. The D127V mutation was localized in the electrostatic belt of the bundle of cytoplasmic helices, indicating that stability of the pentameric bundle of the cytoplasmic helix affects MscL structure. Together, this study described a novel class of MscL mutations that were correlated with the thermodynamic stability of the MscL structure.

fusB is an allele of nadD, encoding nicotinate mononucleotide adenylyltransferase in Escherichia coli

Microbiology ◽  
2003 ◽  
Vol 149 (9) ◽  
pp. 2427-2433 ◽  
Author(s):  
Martin Stancek ◽  
Leif A. Isaksson ◽  
Monica Rydén-Aulin
Keyword(s):  
Escherichia Coli ◽  
Amino Acids ◽  
Temperature Sensitivity ◽  
Minimal Medium ◽  
Synthetic Medium ◽  
Normal Growth ◽  
Temperature Sensitive ◽  
Permissive Temperature ◽  
Atp Binding Site ◽  
Pleiotropic Phenotype

Isolation of the temperature-sensitive Escherichia coli mutant 72c has been described previously. The mutant allele was named fusB and causes a pleiotropic phenotype, the most striking features of which, besides temperature sensitivity, are the inability to grow on synthetic medium and supersensitivity to trimethoprim, an antibiotic that inhibits the C1 metabolism. This work shows that the fusB mutation is a frameshift mutation in the nadD gene that encodes nicotinate mononucleotide adenylyltransferase. The frameshift leads to a change of the last 10 amino acids and an addition of 17 amino acids. This lesion, renamed nadD72, leads to very little NAD+ and NADPH synthesis at the permissive temperature and essentially no synthesis at the non-permissive temperature. As a comparison, a new mutation in the nadD gene, with an amino acid change in the ATP-binding site, has been isolated. Its NAD+ synthesis is decreased at 30 °C but the level is still sufficient to support normal growth. At 42 °C, NAD+ synthesis is reduced further, which leads to temperature sensitivity on minimal medium. This mutation was designated nadD74. Thus, a small decrease in NAD+ levels affects ability to grow on minimal medium at 42 °C, while a large decrease leads to a more pleiotropic phenotype.

scholarly journals TEMPERATURE-SENSITIVE MUTANTS FOR THE REPLICATION OF PLASMIDS IN ESCHERICHIA COLI II. PROPERTIES OF HOST AND PLASMID MUTATIONS

Genetics ◽  
1973 ◽  
Vol 74 (1) ◽  
pp. 1-16
Author(s):  
David T Kingsbury ◽  
Donna G Sieckmann ◽  
Donald R Helinski
Keyword(s):  
Escherichia Coli ◽  
Dna Replication ◽  
Cell Growth ◽  
Dna Synthesis ◽  
Plasmid Dna ◽  
Recipient Cell ◽  
Conjugal Transfer ◽  
Temperature Sensitive ◽  
Permissive Temperature ◽  
Group I

ABSTRACT Host mutations in Escherichia coli K12 selected for the temperature-sensitive replication of the bacterial plasmid colicinogenic factor E1 (ColE1) exhibit a pleiotropic effect with respect to the effect of the mutation on other extrachromosomal elements. The mutations also vary with respect to the time of incubation of the cells at 43°C required for complete cessation of COlE1, DNA synthesis. While the synthesis of the bacterial chromosome appears unaffected, supercoiled ColE1 DNA replication stops immediately in some mutants and gradually decreases during several generations of cell growth before stopping in others. Mutations isolated in the ColE1 plasmid resulted in only a gradual cessation of ColE1 DNA synthesis over several generations of cell growth at 43°C. Conjugal transfer of the ColE1 and COlV factors occurs normally in the host mutants when the transfer is carried out at the permissive temperature; however, the presence of a group I mutation in the donor cell prohibited conjugal transfer of either plasmid DNA at 43°C to a normal recipient cell. Similarly, the presence of this mutation in the recipient prevented the establishment of COlE1 or COlV in the mutant recipient cell upon conjugation with a normal donor at 43°C. Various host COlE1, replication mutants carrying either ColE1 or ColE2 were also defective in the mitomycin Cinduced production of colicin E1 or colicin E2 at 43°C. The majority of the host mutations examined exhibited a temperature sensitivity to growth in deoxycholate in addition to the inhibition of plasmid DNA replication, suggesting a membrane alteration in these mutants when grown at the restrictive temperature.

scholarly journals tRNA2Thr Complements Temperature Sensitivity Caused by Null Mutations in the htrB Gene in Escherichia coli

2003 ◽  
Vol 185 (5) ◽  
pp. 1726-1729 ◽  
Author(s):  
Yoshio Mohri ◽  
Simon Goto ◽  
Kenji Nakahigashi ◽  
Hachiro Inokuchi
Keyword(s):  
Escherichia Coli ◽  
Protein Synthesis ◽  
Temperature Sensitivity ◽  
Lipid A ◽  
Temperature Sensitive ◽  
Temperature Sensitive Mutants ◽  
Lipid A Biosynthesis ◽  
Two Temperature ◽  
Temperature Sensitive Phenotype

ABSTRACT According to the wobble rule, tRNA2Thr is nonessential for protein synthesis, because the codon (ACG) that is recognized by tRNA2Thr is also recognized by tRNA4Thr. In order to investigate the reason that this nonessential tRNA nevertheless exists in Escherichia coli, we attempted to isolate tRNA2Thr-requiring mutants. Using strain JM101F−, which lacks the gene for tRNA2Thr, we succeeded in isolating two temperature-sensitive mutants whose temperature sensitivity was complemented by introduction of the gene for tRNA2Thr. These mutants had a mutation in the htrB gene, whose product is an enzyme involved in lipid A biosynthesis. Although it is known that some null mutations in the htrB gene give a temperature-sensitive phenotype, our mutants exhibited tighter temperature sensitivity. We discuss a possible mechanism for the requirement for tRNA2Thr.

Suppression of the ssb-1 and ssb-113 mutations of Escherichia coli by a wild-type rep gene, NaCl, and glucose

1982 ◽  
Vol 152 (2) ◽  
pp. 572-583
Author(s):  
E S Tessman ◽  
P K Peterson
Keyword(s):  
Escherichia Coli ◽  
Temperature Sensitivity ◽  
Genetic Background ◽  
Growth Defect ◽  
Temperature Sensitive ◽  
Wild Type ◽  
Strong Suppression ◽  
Uv Sensitivity ◽  
Ssb Proteins ◽  
K 12

The ssb-1 mutation confers severe temperature sensitivity and UV sensitivity on many strains of Escherichia coli K-12 and C, including strain C1412. However, ssb-1 confers only slight temperature sensitivity and slight UV sensitivity on strain C1a, suggesting that strain C1a contains extragenic suppressors of ssb-1. We found that introduction of the wild-type rep gene from C1a into strain C1412 ssb-1 gave strong suppression of temperature sensitivity and moderate suppression of UV sensitivity. Also, the C1a rep+ gene mildly suppressed the temperature sensitivity conferred by the ssb-113 mutation, formerly called lexC113. Suppression of the C1412 ssb-1 growth defect by C1a rep+ rendered the cells Gro- for phi X174. In contrast to the positive suppression of ssb-1 and ssb-113 by a wild-type rep gene, mutant rep alleles enhanced the severity of the ssb-1 defect, with several C1a ssb-1 double mutants being either more temperature sensitive or more UV sensitive than C1a ssb-1, depending on which mutant rep allele was used. As a control, the same rep alleles in combination with a dnaB mutation gave an allele-independent increase in temperature sensitivity. Our results on suppression of ssb-1 by rep and on the role of the genetic background in this suppression suggested that the rep and ssb proteins interact to form a subcomplex of the total DNA replication complex and that this subcomplex has some function in repair. The effects of NaCl and glucose on suppression of both the temperature sensitivity and the UV sensitivity conferred by ssb-1 and ssb-113 are described. The degree of suppression of temperature sensitivity by salt or glucose was dependent on the source of the wild-type rep allele, as well as on the genetic background.

scholarly journals ISOLATION, GENETIC MAPPING AND SOME CHARACTERIZATION OF A MUTATION IN ESCHERICHIA COLI THAT AFFECTS THE PROCESSING OF RIBONUCLEIC ACID

Genetics ◽  
1978 ◽  
Vol 90 (4) ◽  
pp. 659-671
Author(s):  
David Apirion
Keyword(s):  
Escherichia Coli ◽  
Temperature Sensitivity ◽  
Single Point ◽  
5S Rrna ◽  
23S Rrna ◽  
Single Point Mutation ◽  
Temperature Sensitive ◽  
Rnase E ◽  
Nonpermissive Temperature ◽  
Ribonuclease E

ABSTRACT Temperature-sensitive mutants were isolated from an rnc (RNase III-) strain of Escherichia coli, and their rRNA metabolism was analyzed on 3% polyacrylamide gels. One of these mutants was unable to produce 23S and 5S rRNAs at the nonpermissive temperature. When an rnc  + allele was introduced to this strain, it remained temperature sensitive. At the nonpermissive temperature, this strain could then produce 23S rRNA but was unable to make normal levels of 5S rRNA. In matings and transduction experiments, the defect in rRNA metabolism and temperature sensitivity behaved as a syndrome caused by a single point mutation, which was mapped at min 23.5 on the E. coli chromosome. This mutation probably affects an enzyme, ribonuclease E (RNase E), which introduces a cut in the nascent rRNA transcript between the 23S and the 5S rRNA cistrons. The mutation rne is recessive with respect to temperature sensitivity and the pattern of rRNA. Revertants able to grow at 43° and with normal metabolism of rRNA were isolated; genetic analysis showed that they do not contain the original rne mutation, suggesting that they were true revertants. By combining the rne mutation with an rnc mutation, double rnc rne strains were synthesized, which behaved very similarly to the original rnc strain from which the rne mutation was isolated. Such strains have RNA metabolism that is similar to that of rnc strains at permissive temperatures, but at the nonpermissive temperature they fail to synthesize p23, m23 and 5S rRNAs. Thus, the experiments reported here, together with previous studies, suggest the existence of a new processing ribonuclease activity in Escherichia coli, which is called ribonuclease E.

scholarly journals Temperature Sensitivity of Bacteriolysis Induced by β-Lactam Antibiotics in Amino Acid-Deprived Escherichia coli

1998 ◽  
Vol 180 (8) ◽  
pp. 2224-2227 ◽  
Author(s):  
Dmitrii G. Rodionov ◽  
Edward E. Ishiguro
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Heat Shock ◽  
Temperature Sensitivity ◽  
Temperature Sensitive ◽  
Shock Response ◽  
Tolerance Response ◽  
Shock Proteins ◽  
Temperature Upshift ◽  
Induction Stage

ABSTRACT The temperature-sensitive penicillin tolerance response previously reported in amino acid-deprived Escherichia coli (W. Kusser and E. E. Ishiguro, J. Bacteriol. 169:2310–2312, 1987) was not due to the induction of the heat shock response resulting from a temperature upshift and was therefore unrelated to the findings of another report (J. K. Powell and K. D. Young, J. Bacteriol. 173:4021–4026, 1991) indicating a positive correlation between the expression of heat shock proteins and penicillin tolerance. The thermosensitive event occurred in the lysis induction stage.

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