Sequence of the T4 recombination gene,uvsX, and its comparison with that of therecAgene ofEscherichia coil

Abstract

We have determined the nucleotide sequence of the uvsX gene of bacteriophage T4 which is involved in DNA recombination and damage repair, and whose product catalyzes in vitro reactions related to recombination process in analogous manners to E. coli recA gene product. The coding region consisted of 1170 nucleotides directing the synthesis of a polypeptide of 390 amino acids in length with a calculated molecular weight of 43,760. Amino acid composition, the sequence of seven NH2-terminal amino acids and molecular weight of the protein deduced from the nucleotide sequence were consistent with the data from the analysis of the purified uvsX protein. The nucleotide sequence and the deduced amino acid sequence were compared with those of the recA gene. Although a significant homology was not found in the nucleotide sequences, the amino acid sequences included 23% of identical and 15% of conservatively substituted residues.

Homologous genetic recombination as an intrinsic dynamic property of a DNA structure induced by RecA/Rad51-family proteins: A possible advantage of DNA over RNA as genomic material

“…Some of the RecA͞Rad51-family proteins were demonstrated by electron microscopy to form conserved, righthanded, spiral filamentous structures either by themselves or around DNA that is either single-stranded or double-stranded (20,21), and to some extent to have conserved amino acid sequences within the core region. It is remarkable that UvsX is only slightly conserved in amino acid sequence (22), but is well conserved in its biochemical functions and three-dimensional structure (16,23,24). The three-dimensional structure of DNAfree RecA, as solved by x-ray crystallography, revealed a righthanded helical filament (25), which is consistent with that obtained by electron microscopy as described above.…”

Domain structure and dynamics in the helical filaments formed by RecA and Rad51 on DNA

“…Both ADP-AlF 4 Ϫ and ATP-␥-S are being used in this case as analogs for ATP, but it can clearly be seen that the filament structure depends on which analog is being used. Numerous studies have shown that aluminum fluoride can serve as a very good non-hydrolyzable analog for ATP (28), whereas it has been shown that ATP-␥-S is slowly hydrolyzed by proteins such as RecA (29)(30)(31). The smaller pitch filaments, resembling the ''collapsed'' state of the RecA filament formed in the absence of ATP (25), have previously been seen with hRad51 protein on ssDNA when ATP-␥-S was used as a cofactor (23).…”

A role of RnlA in the RNase LS activity from Escherichia coli

“…The amplified DNA fragment was digested with EcoRI and cloned downstream of a λ phage promoter, P L , in the plasmid pNT45 (Fujisawa et al, 1985) to construct pGE615. pETN expressing RnlA was constructed as follows.…”

“…Purification of Dmd MH1 cells were co-transformed with pGE615 and pNT204 (Fujisawa et al, 1985), the latter of which encodes a temperature-sensitive λ CI repressor. Transformed cells were incubated at 30°C to a density of 2 × 10 8 cells/ml in 500 ml LB medium, and the culture was shifted to 42°C for 2 hr to induce Dmd protein.…”