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Identification of four acidic amino acids that constitute the catalytic center of the RuvC Holliday junction resolvase

, : Identification of four acidic amino acids that constitute the catalytic center of the RuvC Holliday junction resolvase. Proceedings of the National Academy of Sciences of the United States of America 92(16): 7470-7474

Escherichia coli RuvC protein is a specific endonuclease that resolves Holliday junctions during homologous recombination. Since the endonucleolytic activity of RuvC requires a divalent cation and since 3 or 4 acidic residues constitute the catalytic centers of several nucleases that require a divalent cation for the catalytic activity, we examined whether any of the acidic residues of RuvC were required for the nucleolytic activity. By site-directed mutagenesis, we constructed a series of ruvC mutant genes with similar amino acid replacements in 1 of the 13 acidic residues. Among them, the mutant genes with an alteration at Asp-7, Glu-66, Asp-138, or Asp-141 could not complement UV sensitivity of a ruvC deletion strain, and the multicopy mutant genes showed a dominant negative phenotype when introduced into a wild-type strain. The products of these mutant genes were purified and their biochemical properties were studied. All of them retained the ability to form a dimer and to bind specifically to a synthetic Holliday junction. However, they showed no, or extremely reduced, endonuclease activity specific for the junction. These 4 acidic residues, which are dispersed in the primary sequence, are located in close proximity at the bottom of the putative DNA binding cleft in the three-dimensional structure. From these results, we propose that these 4 acidic residues constitute the catalytic center for the Holliday junction resolvase and that some of them play a role in coordinating a divalent metal ion in the active center.


PMID: 7638215

DOI: 10.2307/2368266

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Ofuji, M.; Ota, Z., 1977: Pyrexia, arthralgia and a consciousness disorder: (systemic lupus erythematosus and systemic aspergillosis). Nihon Rinsho. Japanese Journal of Clinical Medicine 35 Suppl 2: 3020-1, 3382-3

Manter, H.W., 1970: A new species of Transversotrema (Trematoda: Digenea) from marine fishes of Australia. Transversotrema licinum n.sp. is described from Scorpius sp. and Microcanthus strigatus in Queens-land, Australia. It is ectoparasitic beneath the scales and is regarded as a normal adult, not a progenetic metacercaria. It differs from related spe...

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Eide, T.; Marsden, H.S.; Leib, D.A.; Cunningham, C.; Davison, A.J.; Langeland, N.; Haarr, L., 1999: Identification of the UL4 protein of herpes simplex virus type 1. Herpes simplex virus type 1 gene UL4 is predicted to encode a 199-amino-acid protein with a molecular mass of 21 5 kDa. We report here identification of this protein and its localization in the nuclei of infected cells. Antisera raised against oli...

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Shanahan T.G.; Mehta M.P.; Gehring M.A.; Buchler D.A.; Kubsad S.S.; Kinsella T.J., 1989: Minimization of small bowel volume utilizing customized belly board mold. International Journal of Radiation Oncology, Biology, Physics 17(SUPPL 1): 187-188

Penneau, M.; Desnos, J., 1977: Ossifying fibroma of the mandible. Journal Francais D'oto-Rhino-Laryngologie; Audiophonologie, Chirurgie Maxillo-Faciale 26(8): 631-633

Zhou J.; Shen J.; Liu M., 1983: A cytological study on the gynogenesis of fang zheng crucian carp of heilongjiang province china. Gynogenesis was studied in Fang-Zheng Crucian carp. Three groups of gynogenetic processes, including Crucian carp (.female.) .times. common carp (.male.), have been observed. During fertilization, the sperm, either of the same species or of the in...

Quraishi, A.; Rossignol Bancilhon, L.; Nozeran, R., 1979: Effect of the origin of the explant on callus formation and organogenesis in vitro of Solanum tuberosum L. cv. BF 15. Callus formation and organogenesis were more successful in cultures of stems from tuber sprouts than from the sprouts themselves and in nodal cuttings without buds rather than internodal cuttings and from stem fragments rather than tuber fragments...