Substrate Specificity of Nuclease P1

Fujimoto, Masao; Kuninaka, Akira; Yoshino, Hiroshi

doi:10.1271/bbb1961.38.1555

Cited by 121 publications

(63 citation statements)

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“…The incubation mixture contained 5'-32P or 3'-32P end-group-labelled RNA, carrier yeast tRNA and nuclease PI [41] at a ratio of 15 ng/100 pg of RNA. Incubation was in 50 mM ammonium acetate buffer pH 5.3, at 20°C.…”

Section: Partial Digestion Of End-group-labelled Rna With Nuclease Pimentioning

confidence: 99%

Studies on the Sequence of the 3′‐Terminal Region of Turnip‐Yellow‐Mosaic‐Virus RNA

Silberklang

Prochiantz

Haenni

et al. 1977

European Journal of Biochemistry

193

107

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A fragment representing the 3′‐terminal ‘tRNA‐like’ region of turnip yellow mosaic (TYM) virus RNA has been purified following incubation of intact TYM virus RNA with Escherichia coli‘RNase P’. This fragment, which is 112 ± 3‐nucleotides long has been completely digested with T1 RNase and pancreatic RNase and all the oligonucleotides present in such digests have been sequenced using 32P‐end labelling techniques in vitro. The TYM virus RNA fragment is free of modified nucleosides and does not contain a G‐U‐U‐C‐R sequence. Using nuclease P1 from Penicillium citrinum, the sequence of 26 nucleotides from the 5′ end and 16 nucleotides from the 3′ end of this fragment has been deduced. The nucleotide sequence at the 5′ end of the TYM virus RNA fragment indicates that this fragment includes the end of the TYM virus coat protein gene.

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Section: Partial Digestion Of End-group-labelled Rna With Nuclease Pimentioning

confidence: 99%

Studies on the Sequence of the 3′‐Terminal Region of Turnip‐Yellow‐Mosaic‐Virus RNA

Silberklang

Prochiantz

Haenni

et al. 1977

European Journal of Biochemistry

193

107

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“…In contrast, P1 nuclease possesses no pyrophosphatase activity, thus rendering cap structures impervious to its hydrolytic activity. Instead, the enzyme is a phosphodiesterase that cleaves the phosphate bond between nucleosides in mRNA, generating 5Ј-nucleotide monophosphates, and releasing the unhydrolyzed 5Ј-terminal cap structures (11,12). TAP and P1 nuclease digests of the radiolabeled mRNA were separated by TLC and analyzed by phosphorimaging (Fig.…”

Section: Resultsmentioning

confidence: 99%

Deletion of Individual mRNA Capping Genes Is Unexpectedly Not Lethal to Candida albicans and Results in Modified mRNA Cap Structures

Dunyak¹,

Everdeen²,

Albanese³

et al. 2002

Eukaryot Cell

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Eukaryotic mRNAs are modified at the 5 end with a cap structure. In fungal cells, the formation of the mRNA cap structure is catalyzed by three enzymes: triphosphatase, guanylyltransferase, and methyltransferase. Fungal capping enzymes have been proposed to be good antifungal targets because they differ significantly from their human counterparts and the genes encoding these enzymes are essential in Saccharomyces cerevisiae. In the present study, Candida albicans null mutants were constructed for both the mRNA triphosphatase-encoding gene (CET1) and the mRNA methyltransferase encoding gene (CCM1), proving that these genes are not essential in C. albicans. Heterozygous deletions were generated, but no null mutants were isolated for the guanylyltransferase-encoding gene (CGT1), indicating that this gene probably is essential in C. albicans. Whereas these results indicate that Cet1p and Ccm1p are not ideal molecular targets for development of anticandidal drugs, they do raise questions about the capping of mRNA and translation initiation in this fungus. Southern blot analysis of genomic DNA indicates that there are not redundant genes for CET1 and CCM1 and analysis of mRNA cap structures indicate there are not alternative pathways compensating for the function of CET1 or CCM1 in the null mutants. Instead, it appears that C. albicans can survive with modified mRNA cap structures.

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“…Owing to its very high preference for singlestranded polynucleotides, rye germ nuclease may be classified as a 'single-strand-specific' nuclease resembling those isolated from Neurospora creassa [6,7], Aspergillus oryzae [l -51, Penicillum citrinum [39], mung bean [8,9] and wheat seedlings [lo-121. Native double-stranded phage T7 DNA and the helical complex formed between poly(U) and poly(A) are resistant to the action of rye germ nuclease.…”

Section: Discussionmentioning

confidence: 99%

Single‐Strand‐Specific Nuclease from the Nucleoplasm of Rye Germ Nuclei

Przykorska

Szarkowski

1980

European Journal of Biochemistry

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1. The localization of DNAase activity in rye germ nuclei revealed 48% of the total nuclear activity in the nucleoplasm and 4 % in the chromatin.2. The endonuclease present in the nucleoplasm was purified more than 300-fold as compared with the specific activity in homogenized nuclei by means of ammonium sulphate fractionation and CM-cellulose and CM-Sephadex column chromatography. The preparation gave a single band on polyacrylamide gels. Its molecular weight is 45000, pH optimum 5.4. The preparation does not exhibit activity of non-specific phosphomonoesterase and phosphodiesterase.3 . The nuclease catalyses the hydrolysis of denatured DNA and RNA to acid-soluble 5'-phosphate-terminated products and cleaves the phosphomonoester linkage of 3'AMP.4. Synthetic polyribonucleotides are hydrolysed at a rate decreasing in the order : poly(U) The endonuclease shows a high preference for single-stranded nucleic acids. It does not depolymerise double-stranded poly(U) . poly(A).6. The nuclease attacks covalently closed circular bacteriophage PM2 DNA, plasmid ColEl and plasmid pBR322 DNA, converting it initially into the open-circular (relaxed) form and subsequently into the linear form. This reaction is strongly dependent on ionic strength.7. The enzyme nicks the supercoiled DNA molecule of phage PM2 in only one place. At high enzyme concentrations several nicks in such a molecule are observed.

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Substrate Specificity of Nuclease P1

Cited by 121 publications

References 0 publications

Studies on the Sequence of the 3′‐Terminal Region of Turnip‐Yellow‐Mosaic‐Virus RNA

Studies on the Sequence of the 3′‐Terminal Region of Turnip‐Yellow‐Mosaic‐Virus RNA

Deletion of Individual mRNA Capping Genes Is Unexpectedly Not Lethal to Candida albicans and Results in Modified mRNA Cap Structures

Single‐Strand‐Specific Nuclease from the Nucleoplasm of Rye Germ Nuclei

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