Toshiaki Kayano

Nucleotide sequence of the genes for β and ε subunits of proton-translocating ATPase from Escherichia coli

Abstract The 1855-nucleotide long DNA sequence of part of the gene cluster for the proton-translocating ATPase from E. coli was determined by the method of Maxam-Gilbert. The sequence covers the genes for the β and e subunits of F1 along with the flanking region. The amino acid sequence of these subunits deduced from the nucleotide sequence indicates that the β and e subunits have 459 and 138 amino acids, respectively. The possible secondary structure of the both subunits was estimated from the deduced primary structures. A possible nucleotide binding site in the β subunit is also discussed on the basis of the primary and secondary structures. The codons used in the genes for all the components of F1F0 were different in different genes, suggesting that the amount of each subunit in the F1F0 is determined to some extent on a translational level.

Nucleotide sequence of the genes coding for α, β and γ subunits of the proton-translocating ATPase of Escherichia coli

Abstract A nucleotide sequence of 2328 base pairs comprising a portion of the gene cluster for the proton-translocating ATPase of E. coli was determined. The sequence covers most of the gene for α subunit, the entire gene for γ subunit and the amino terminal portion of the gene for β subunit, along with the flanking regions of these genes. The amino acid sequences of these subunits deduced from the DNA sequences indicate that the α and γ subunits have 513 and 287 amino acid residues, respectively. A possible secondary structure for each subunit was estimated from the inferred primary structure. The intercistronic regions between the genes for α and γ and between γ and β are 49 and 26 base pairs, respectively. The significance of codon usage in these genes is discussed in correlation with their expression.

Nucleotide sequence of the genes for F0 components of the proton-translocating ATPase from Escherichia coli: Prediction of the primary structure of F0 subunits

The 1763 nucleotide-long-DNA sequence of part of the gene cluster for the proton-translocating ATPase from E. coli was determined. The sequence covers the genes for the a and b subunits of F0 along with the intercistronic regions. In the region preceding the gene for the a subunit, a reading frame encompassing 127 amino acids was found. The primary structure of the a and b subunits were deduced and the properties of these proteins were predicted. Analysis of codon usage in these genes was made.

Nucleotide sequence of genes coding for dicyclohexylcarbodiimide-binding protein and the α subunit of proton-translocating ATPase of Escherichia coli

Summary The recombinant plasmid pMCR533 carries part of the gene cluster coding for the proton-translocating ATPase (F1-F0) of E. coli. The DNA sequence of the structural gene for dicyclohexylcarbodiimide-binding protein was obtained by analyzing a DNA fragment from this plasmid. The DNA sequence corresponding to the amino terminus of the structural gene for the α subunit was also found on the plasmid. Both sequences were in good agreement with the amino acid sequences determined previously. From the exact locations of these two genes we have located genes coding for other components of F1-F0 on the defined portion of E. coli DNA. A proposal is made to use the nomenclature pap (proton-translocating ATPase subunit protein) for the structural gene cluster for F1-F0.

Nucleotide sequence of the gene coding for the δ subunit of proton-translocating ATPase of Escherichia coli

Abstract The DNA sequence of a part of the gene cluster for the proton-translocating ATPase of E. coli was determined. Two reading frames were found between the genes for dicyclohexylcarbodiimide-binding protein and α subunit. The following evidence indicates that one frame codes the δ subunit; i) The primary structure deduced from DNA sequence agreed with amino acid composition of the protein. ii) Five residues from the amino terminal were the same as those deduced from DNA sequence. The α helix content of this protein (estimated from the primary structure) was 60.2% of the total residues with the longest helical domain of 50 residues. The intercistronic sequence between the genes for δ and α had 15 base pairs, suggesting that the synthesis of the α is not regulated transcriptionally. The organization of the gene cluster was also shown on the physical map.

Isolation and characterization of stress-responsive genes from the scleractinian coral Pocillopora damicornis

In order to identify the expression of coral genes induced by stressors such as suspended red soil, mRNA differential display was examined for the coral Pocillopora damicornis. Apparent differences were observed in the gene expression profiles between control colonies and those treated with local red soil, indicating that red soil affects gene expression in P. damicornis. We cloned nine candidate PCR fragments derived from the differentially expressed genes. One of the clones, pPd9-1, showed a high similarity to a member of the heat-shock protein (HSP) 70 family, suggesting that red soil may cause protein denaturation in the coral. Reverse transcription-mediated PCR showed that the expression of pPd9-1 was also increased by elevated temperature, but not by reduced salinity.

Evolutionary origin of two genes for chloroplast small heat shock protein of tobacco

Two different cDNA clones for the chloroplast small heat shock protein (smHSP) were isolated from tobacco (Nicotiana tabacum cv. Petit Havana SR1). One of the cDNAs (type I) has a full-length open reading frame (ORF) of the smHSP of 26.6 kDa. By contrast, the other one (type II) contains an additional nucleotide that causes the frame shift inside a putative ORF for the smHSP. If this nucleotide is neglected, type II cDNA encodes the smHSP that is 89% identical to that encoded by type I cDNA. Southern blot and polymerase chain reaction (PCR) analyses with genomic DNA indicated that tobacco has two different smHSP genes while two ancestors of tobacco, N. sylvestris and N. tomentosiformis, have a single gene that each corresponds to one of the two genes of tobacco. It was also found that one of the tobacco genes has an ORF for the smHSP disrupted by nucleotide insertion in the same way as type II cDNA, while both ancestor genes have a functional ORF. These results suggest that the two smHSP genes of tobacco had been derived from the two ancestor species, and that one of the two genes had been disrupted by nucleotide insertion during the course of the evolution of tobacco. Northern blot and reverse transcription (RT)-PCR analyses demonstrated that both the tobacco genes are expressed upon heat stress, exhibiting different dependences on temperature.