The structural and sequence homology of a family of microbial ribonucleases

Hill, C.P.; Dodson, Guy; Heinemann, Udo; Saenger, Wolfram; Mitsui, Yukio; Nakamura, Kazuo; Борисов, С. А.; Tischenko, Galina; Polyakov, K. M.; Pavlovsky, Sasha

doi:10.1016/0968-0004(83)90363-8

Cited by 146 publications

(89 citation statements)

References 23 publications

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“…Asn39 appears important in maintaining the conformation around the binding site for the nucleotide. The side chains of Glu54, Arg69 and His85 are proposed as catalytic residues (Hill et al, 1983).…”

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confidence: 99%

Complex of ribonuclease Sa with a cyclic nucleotide and a proposed model for the reaction intermediate

Ševčı́k¹,

Zegers²,

Wyns³

et al. 1993

European Journal of Biochemistry

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The structure of the complex of ribonuclease from Streptomyces aureofaciens (RNase Sa) with ex0 guanosine 2',3'-cyclophosphorothioate has been refined against 0.2-nm resolution synchrotron data using, as a starting model, coordinates from the RNase Sa: 2'-GMP complex. The refinement was based on all data over 1.0-0.2 nm and converged to a crystallographic R factor of 11.9%. This is the first structure of a microbial ribonuclease complexed with a 2',3'-cyclophosphorothioate, which is a thio analogue of the intermediate of the two-step reaction. However, ex0 guanosine 2',3'-cyclophosphorothioate is bound in a non-functional mode and is not hydrolysed. This structure therefore does not provide direct evidence on the identity of the amino acid residues responsible for catalytic cleavage of the substrate. However, based on present and previous results, a plausible model is proposed for the complex of the cyclic intermediate which acts as substrate for the second step of the catalysis.

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confidence: 99%

Complex of ribonuclease Sa with a cyclic nucleotide and a proposed model for the reaction intermediate

Ševčı́k¹,

Zegers²,

Wyns³

et al. 1993

European Journal of Biochemistry

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“…bers of the fungal RNase T1 family, such as RNase T1 of Aspergillus oryzae [20] and RNase N1 of Neurospora crassa [21], showed the following extent of identity. VSTAQAAGYQLHEDGETVGSNSYPHKYNNYEGFDFSVSSPYY * * * * * * * * * * * A. oryzae RNase T1 MSSNLIKSFGLIAIGAISGVTFTHFYYKGYQGSDVPDLTPRY * * * ** * * * * * * * * S. pombe nuclease…”

Section: Resultsmentioning

confidence: 99%

Purification, characterization and complete amino acid sequence of nuclease Cl from Cunninghamella echinulata var. echinulata

Ho¹,

Shiau²,

Liu³

et al. 1998

European Journal of Biochemistry

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It is known, from the zymogram method of nuclease activity assay, that the crude extracts of Cunninghamella echinulata var. echinulata contained at least three distinct extracellular nucleases. Among them, the major form was 30 kDa in molecular mass and termed nuclease C1. In this report, nuclease C1 was purified to apparent homogeneity by chromatography on Cibacron blue-3GA, phenyl-Sepharose 4B and HiTrap Heparin. Nuclease C1 acquired enzymatic activity in the presence of Mn 2ϩ or Mg 2ϩ and was inhibited by EDTA. The activity was maximal at pH 7Ϫ8.5. The primary structure of nuclease C1 was completely determined using enzymatic digestion and gene cloning. The N-terminal 49 residues of nuclease C1 were first elucidated from a tryptic digest. Two degenerate upstream primers were subsequently designed to amplify the cDNA encoding nuclease C1. The resulting protein sequence of nuclease C1 was shown to be composed of 252 residues. It was intriguing to find that the protein sequence of nuclease C1 showed significant similarities with the sequences of the mitochondrial nucleases of Saccharomyces cerevisiae (44% identity) and Schizosaccharomyces pombe (42 % identity). Residue His87 of nuclease C1 was postulated to be located at the active site from sequence similarity with secreted nuclease from Serratia marcescens.Keywords : nuclease C1; mitochondrial nuclease ; Cunninghamella echinulata ; Saccharomyces cerevisiae; Schizosaccharomyces pombe.Nucleases act on both DNA and RNA to release products containing 5′-or 3′-phosphates [1]. Several fungal species are known to produce both extracellular and intracellular nucleases. Representatives of extracellular fungal nucleases are nuclease S1 of Aspergillus oryzae [2,3] and nuclease P1 of Penicillium citrinum [4,5]. The properties and the structures of these two nucleases are well characterized. Other fungi also secrete nucleases, e.g. Neurospora crassa and Aspergillus nidulans, which are referred to as DNase (deoxyribonuclease) A [6] and DNase 4 [7], respectively.Clinically, some Cunninghamella species cause severe syndromes and even led to death in humans. For example, several reports have indicated that Cunninghamella bertholletiae can infect patients with cancer and immunocompromise children [8,9]. Diseases including paranasal sinusitus [10] and mucormycosis [11] have been relatively correlated with Cunninghamella infection. The most important risk factors for C. bertholletiae infection include corticosteroid administration and pro-

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“…Streptomyces aureofaciens bacteria secrete extracellular ribonucleases which are members of the extended microbial family (Hartley, 1980;Hill et al, 1983). RNases have been isolated and characterized from three strains of S. aureofaciens to date and referred to as RNase Sa, Sa2 and Sa3.…”

Section: Introductionmentioning

confidence: 99%

“…The functional role of these enzymes has not been clearly established. Those studied so far all appear to have the same overall fold (Hill et al, 1983;Sev61"k et al, 1990), but there is only low sequence identity between many members of the family, with substantial deletions and insertions in the surface loops. Some of the organisms produce an intracellular polypeptide inhibitor of their ribonuclease to prevent it being active prior to secretion, which would be extremely harmful to the cell.…”

Section: Introductionmentioning

confidence: 99%

Recognition of RNase Sa by the Inhibitor Barstar: Structure of the Complex at 1.7 Å Resolution

et al. 1998

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The 1.7 .~ resolution structure of RNase Sa complexed with the polypeptide inhibitor barstar is reported here. The crystals are in the hexagonal space group~ P65 with unit-cell dimensions a = b = 56.9, c = 135.8 A and the asymmetric unit contains one molecule of the complex. RNase Sa is an extracellular microbial ribonuclease produced by Streptomyces aureofaciens. Barstar is the natural inhibitor of barnase, the ribonuclease of Bacillus amyloliquefaciens. It inhibits RNase Sa and barnase in a similar manner by steric blocking of the active site. The structure of RNase Sa is very similar to that observed in crystals of the native enzyme and its complexes with nucleotides. Barstar retains the structure found in its complex with barnase. The accessible surface area of protein buried in the complex is about 300 ~2 smaller and there are fewer hydrogen bonds in the enzymeinhibitor interface in RNase Sa-barstar than in barnasebarstar, providing an explanation of the reduced binding affinity in the former. Previous studies of barstar complexes have used mutants of the inhibitor and this is the first structure which includes wild-type barstar.

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The structural and sequence homology of a family of microbial ribonucleases

Cited by 146 publications

References 23 publications

Complex of ribonuclease Sa with a cyclic nucleotide and a proposed model for the reaction intermediate

Complex of ribonuclease Sa with a cyclic nucleotide and a proposed model for the reaction intermediate

Purification, characterization and complete amino acid sequence of nuclease Cl from Cunninghamella echinulata var. echinulata

Recognition of RNase Sa by the Inhibitor Barstar: Structure of the Complex at 1.7 Å Resolution

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