In previous work (E. S. Tessman and P. K. Peterson, J. Bacteriol. 163:677-687 and 688-695, 1985), we isolated many novel protease-constitutive (Prtc) recA mutants, i.e., mutants in which the RecA protein was always in the protease state without the usual need for DNA damage to activate it. Most P&tC mutants were recombinase positive and were designated PrtC Rec+; only a few Prtc mutants were recombinase negative, and those were designated Prtc Rec-. We report changes in DNA sequence of the recA gene for several of these mutants. The mutational changes clustered at three regions on the linear RecA polypeptide. Region 1 includes amino acid residues 25 through 39, region 2 includes amino acid residues 157 through 184, and region 3 includes amino acid residues 298 through 301. The in vivo response of these Prtc mutants to different effectors suggests that the RecA effector-binding sites have been altered. In particular we propose that the mutations may define single-stranded DNA-and nucleoside triphosphate-binding domains of RecA, that polypeptide regions 1 and 3 comprise part of the single-stranded DNA-binding domain, and that polypeptide regions 2 and 3 comprise part of the nucleoside triphosphate-binding domain. The overlapping of single-stranded DNA-and nucleoside triphosphate-binding domains in region 3 can explain previously known complex allosteric effects. Each of four Prtc Rec-mutants sequenced was found to contain a single amino acid change, showing that the change of just one amino acid can affect both the protease and recombinase activities and indicating that the functional domains for these two activities of RecA overlap. A recA promoter-down mutation was isolated by its ability to suppress the RecA protease activity of one of our strong PrtC mutants.

ABSTRACT. Early rehabilitative therapy is important for patients with hypertensive cerebral hemorrhage to improve long-term function of the extremities. Vascular endothelial growth factor (VEGF) is closely associated with the pathogenesis of hypertension. To identify the markers contributing to the genetic susceptibility to hypertensive cerebellar hemorrhage (HCH) and rehabilitative treatment, we examined the potential association between HCH and 12 single nucleotide polymorphisms of the VEGF gene. Participants included 244 patients with HCH and 251 healthy controls from our rehabilitation department. The T allelic frequencies of the rs3025020 (intron 6) and rs3025039 (3'-UTR) polymorphisms were significantly higher in the patients with HCH than in the healthy controls (rs3025020 T allele: P = 0.0002, OR = 1.619, 95%CI = 1.256-2.088; rs3025039 T allele: P = 0.001, OR = 1.682, 95%CI = 1.246-2.270). Strong linkage disequilibrium was observed in three blocks (D' > 0.9), and significantly more C-G-C (rs3025020, rs3025030, and rs3025039) haplotypes (P = 0.001) were found in the controls in block 3. Significantly more T-G-C haplotypes were found in the patients with HCH (P = 0.046). Further genotype and clinical phenotype correlation study of the rs3025039 carriers showed that Fugl-Meyer and Barthel index scores were lower in the patients with the TT genotype relative to CT + CC genotypes (P < 0.01). These findings point to a role for VEGF polymorphism in HCH, and may be informative for future investigations on the pathogenesis of rehabilitative treatment.