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F Kawamura

Publications and source records attributed to F Kawamura.

At least 55 records · Page 3Linked to original sources
PubMed1990

[Investigation of a patient with difficult endotracheal intubation].

Unusually difficult endotracheal intubation was encountered during anesthetic induction of a 41-year-old male, necessitating use of a bronchofiberscope. The usual pre-operative analysis had shown no problems, such as tracheal tumor or stricture. Due to difficult intubation, post-operative evaluations were made and no abnormalities were found. However, investigation of the lateral roentgenogram showed a much greater than average backward bend in the patient's trachea under the vocal cord. It is likely that the sharp bend of the trachea was responsible for difficult intubation.

Adult↗
PubMed1989

Dissection of the expression signals of the spoA gene of Bacillus subtilis: glucose represses sporulation-specific expression.

The expression of the spo0A-lacZ fusion gene was partially repressed in the presence of an excess of glucose. Expression was restored either by the mutation sigA47(crsA47) or by addition of decoyinine, an inhibitor of GMP synthetase, to the medium. By constructing a lacZ fusion with a smaller fragment of the spo)A gene, we observed a beta-galactosidase profile in which expression was completely repressed by an excess of glucose. This expression was restored by the addition of decoyinine. These results indicate that the expression of the spo0A gene is regulated by at least two different mechanisms, one sensitive to glucose, the other not. Furthermore, the glucose-sensitive regulation was shown to reside at the transcriptional level. It is likely that the reduced expression of the spo0A gene in the presence of glucose at an early stage of sporulation causes the repression of sporulation.

Adenosine↗
PubMed1988

New suppressor mutation sur0B of spo0B and spo0F mutations in Bacillus subtilis.

Two extragenic suppressor mutations, sur0B20 and sur0F1, which restore the sporulation of spo0B or spo0F mutants of Bacillus subtilis to the wild-type level, were obtained. These suppressor mutations were located in the spo0A gene. Their location is close to that of the sof-1 mutation, which suppresses spo0B, spo0E and spo0F mutations. However, spo0 strains bearing the sur0B20 mutation differed in several phenotypic characteristics from spo0 mutants bearing the sof-1 suppressor. Nucleotide sequence analysis revealed that the sur0B20 and sur0F1 mutations resulted in Glu14 to Val and Asn12 to Lys conversion, respectively, in the spo0A gene. This result indicates that sur0B20 is a new suppressor of spo0b and spo0F mutations, whereas sur0F1 is identical to sof-1.

Alleles↗
PubMed1986

Revised assignment for the Bacillus subtilis spo0F gene and its homology with spo0A and with two Escherichia coli genes.

The nucleotide sequences of spo0F mutant genes which block the early sporulation process of Bacillus subtilis were determined. The mutation sites together with the results of complementation tests suggested that an open reading frame for a polypeptide of Mr = 14,229 is the spo0F gene. The deduced amino acid sequence shows striking homology with that of the spo0A gene. In addition, the upstream region involving the rib some binding site of the spo0F coding region is also similar to those of spo0A and spo0B. These homologies suggest that all three genes have a similar function in regulating the initiation of sporulation, and that their expression is controlled by a common mechanism. Clear homology is also seen between the spo0 gene products and the transcriptional control proteins, OmpR and Dye, of Escherichia coli suggesting that the spo0 gene products also are involved in the control of transcription.

Alleles↗
PubMed1986

The effect of spo0 mutations on the expression of spo0A- and spo0F-lacZ fusions.

We have constructed spo0A-lacZ and spo0F-lacZ fusions with a temperate phage vector and have investigated how spo0 gene products are involved in the expression of each of these genes. The expression of spo0A-lacZ and spo0F-lacZ was stimulated at about the time of cessation of vegetative growth in Spo+ cells. This stimulation of spo0A-lacZ was impaired by mutations in the spo0B, D, E, F or H genes but was not affected by mutations in the spo0J or K genes. Similar results were obtained with the spo0F-lacZ fusion. The effect of the spo0A mutation on spo0A-lacZ expression was characteristic: the spo0A-directed beta-galactosidase activity found during vegetative growth was significantly enhanced in the spo0A mutant. This result suggests that spo0A gene expression is auto-regulated being repressed by its own gene product. Another remarkable observation was the effect of the sof-1 mutation, which is known to be a spo0A allele; it suppressed the sporulation deficiency of spo0B, spo0D and spo0F mutants. The spo0A-lacZ stimulation, which is impaired by any one of these spo0 mutations, was restored by the additional sof-1 mutation.

Bacillus subtilis↗
PubMed1986

Use of the Bacillus subtilis subtilisin signal peptide for efficient secretion of TEM beta-lactamase during growth.

We report the development of an efficient Bacillus subtilis secretory system, with the secreted product stably maintained in the medium for 100 h. The system is based on characterization of the subtilisin signal peptidase cleavage site and promoters, catabolite repression of sporulation, presence of a vegetative secreting mechanism, and availability of a protease-deficient strain.

Bacillus subtilis↗
PubMed1985

Catabolite-resistant sporulation (crsA) mutations in the Bacillus subtilis RNA polymerase sigma 43 gene (rpoD) can suppress and be suppressed by mutations in spo0 genes.

The catabolite-resistant sporulation (crsA) mutation is able to overcome the repressive effect of glucose on sporulation in Bacillus subtilis. Three chromosomal crsA mutations, crsA1, crsA4, and crsA47, were transferred by the "gene conversion" process to B. subtilis plasmid pRPD11, which consists of the entire wild-type rpoD coding sequence, encoding the major sigma 43 factor of B. subtilis RNA polymerase, and pUB110. By DNA sequence analysis we showed that all three crsA mutations were identical two-base changes, CCT (proline) to TTT (phenylalanine), within the rpoD coding sequence. Furthermore, the crsA47 mutation restored spo0J and spo0K sporulation to wild-type levels and partially improved the sporulation efficiencies of spo0B, spo0D, and spo0F. Extragenic suppressors (scr) of crsA47 included mutations in spo0A, spo0D, spo0F, and spo0K plus other mutations that have not been specifically identified. Thus sigma 43 appears to be involved in catabolite repression by glucose, to interact either directly or indirectly with spo0 gene products, and to play an important role in the initiation of spore development in B. subtilis.

Bacillus subtilis↗
PubMed1985

Identification of the transcriptional suppressor sof-1 as an alteration in the spo0A protein.

The mutation sof-1 suppresses the sporulation defect of mutations in either the spo0B, spo0E, or spo0F stage 0 sporulation genes. Through the use of integrative plasmids carrying the portion of the chromosome including the spo0A locus and flanking regions, the sof-1 mutation was localized near the spo0A locus. A plasmid carrying a fragment of DNA with sof genetic activity was constructed. Nucleic acid sequence analysis of this fragment revealed a single base change that resulted in a substitution of lysine for asparagine in the 12th codon of the spo0A gene. The results indicate that certain missense mutations in the spo0A gene bypass the necessity for the spo0B, spo0E, and spo0F gene products in sporulation. Several models for the interaction of these gene products may be imagined.

Bacillus subtilis↗
PubMed1985

Translational coupling in Bacillus subtilis of a heterologous Bacillus subtilis-Escherichia coli gene fusion.

Translational coupling was demonstrated in a gene fusion in which the promoter and the N-terminal region of the Bacillus subtilis subtilisin (aprA) gene were fused to a promoterless Tn9-derived chloramphenicol acetyltransferase (CAT; EC 2.3.1.28) gene. Expression of this gene fusion results in the production of a native-sized CAT product, whereas the Tn9-derived CAT gene is usually not translated from its own ribosome binding site in B. subtilis (D. S. Goldfarb, R. L. Rodriguez, and R. H. Doi, Proc. Natl. Acad. Sci. USA 79:5886-5890, 1982). A 178-base-pair deletion, which removed part of the signal peptide and the propeptide of the aprA gene and created a translational stop codon 230 base pairs upstream of the CAT gene ribosome binding site, reduced expression of the CAT gene. A BamHI 10-mer linker insertion into this deletion site, which restored the reading frame and simultaneously removed the translation stop codon, restored CAT gene expression. The data indicate that expression of the CAT gene was dependent on translation of the truncated aprA gene into the ribosome binding site of the CAT gene.

Acetyltransferases↗
PubMed1984

Cloning of sporulation gene spoIIC in Bacillus subtilis.

Specialized transducing phages rho 11spoIIC and phi 105spoIIC, carrying the Bacillus subtilis sporulation gene spoIIC, were constructed by the prophage transformation method. An EcoRI fragment (2.4 MDal) carrying the spoIIC gene was isolated from the phi 105spoIIC genome and recloned into the EcoRI site of plasmid pUB110. The recombinant plasmids corrected the sporulation defect of a Spo- Rec- host, but slightly inhibited the sporulation of a Spo+ Rec- host.

Bacillus subtilis↗
PubMed1984

Construction of a Bacillus subtilis double mutant deficient in extracellular alkaline and neutral proteases.

A mutant strain of Bacillus subtilis carrying lesions in the structural genes for extracellular neutral (nprE) and serine (aprA) proteases was constructed by the gene conversion technique. This mutant had less than 4% of the extracellular protease activity of the wild type and sporulated normally, indicating that neither of these sporulation-associated proteases is essential for development.

Bacillus subtilis↗
PubMed1983

Isolation and mapping of a new suppressor mutation of an early sporulation gene spoOF mutation in Bacillus subtilis.

We constructed an spoOF deletion (spoOF delta S) mutant of Bacillus subtilis by inserting a chromosomal segment carried by plasmid pUBSF delta S. This plasmid carries a 0.7-kilobase pair deletion that removes the spoOF promoter and a part of the structural gene. We used the spoOF deletion mutant to isolate a new intergenic suppressor of the spoOF phenotype, designated sof1. The sof1 suppressor completely restores the sporulation ability of all spoOF defective mutants tested, including spoOF77, spoOF221 and spoOF delta S. The sof1 suppressor maps to the left of lys1 on the B. subtilis chromosome, in a region rich in sporulation markers and distant from the spoOF locus.

Bacillus subtilis↗
PubMed1983

Selection for restriction-induced in vivo deletion in phage vector phi 1E1 of Bacillus subtilis.

Recombinant phage phi 1E1metB, which contains the 4.5-kb EcoRI fragment of Bacillus subtilis DNA, has no HaeIII cleavage sites within the vector phi 1E1 genome but only in the metB insert. When phi 1E1metB was grown in B. subtilis ISR11, which produces BsuR, the isoschizomer of HaeIII, it was restricted and survived with an efficiency of approx. 10(-5). All the survivors were deletion mutants of phi 1E1metB, and only various segments of the insert DNA delineated by HaeIII sites were deleted. The Met+ transforming activities of the DNAs from phi 1E1metB and its deletion derivatives were examined, and the restriction maps of the deletion mutants were correlated with five metB- mutation sites.

Bacillus subtilis↗
PubMed1983

Early sporulation gene spo0F: nucleotide sequence and analysis of gene product.

We have determined the sequence of a 1,162-base-pair DNA fragment containing a spo0F gene which is required for an early stage of sporulation in Bacillus subtilis. The sequence has only one long open reading frame consisting of 173 codons, which has been confirmed to be the spo0F cistron by DNA-mediated transformation and in vitro transcription. In UV-irradiated "maxicells" containing pUBSF13, the plasmid that carries cloned spo0F gene, we have observed the synthesis of a 20-kilodalton polypeptide that is absent from cells carrying a vector plasmid pUB110. The molecular weight of this protein is in agreement with the calculated molecular weight of the spo0F gene product (Mr, 19,065). The putative promoter sequences of spo0F gene were 5' T-A-T-A-A-T 3' at -10 and 5' T-T-G-A-T-T 3' at -35. An octamer sequence, 5' A-A-A-G-G-A-G-G 3', situated 8 base pairs prior to the initiation codon was found to be perfectly complementary with the 3' end of 16S ribosomal RNA. This result offers additional evidence for the proposal by Rabinowitz's group that an extensive mRNA-rRNA interaction is a requirement for efficient translation by B. subtilis ribosomes.

Bacillus subtilis↗
PubMed1981

Cloning of sporulation gene spoOB of Bacillus subtilis and its genetic and biochemical analysis.

A specialized transducing phage carrying a sporulation gene (spoOB) was constructed from Bacillus subtilis temperate phage rho 11 by in vitro and in vivo recombinations. Transformation experiments showed that the spoOB gene resides on a 1.4-megadalton fragment generated by EcoRI endonuclease treatment of the phage deoxyribonucleic acid (DNA). Mutants of this phage which lost transducing activity were isolated and used for genetic complementation tests and the analysis of protein(s) coded by the 1.4-megadalton fragment. The spoOB locus was shown to be composed of one cistron. Sodium dodecyl sulfate-polyacrylamide gel analysis of proteins synthesized in ultraviolet-irradiated cells infected with these phages showed that the 1.4-megadalton fragment codes at least one protein, of molecular weight 39,000, which is synthesized in both vegetative and sporulating cells. A cleavage map of the phage DNA was constructed by use of restriction endonucleases, EcoRI, BamHI, and SalI, and the site of integration of the 1.4-megadalton fragment was determined. Expression and function of the spoOB gene are discussed.

Bacillus subtilis↗