{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2023,3,15]],"date-time":"2023-03-15T21:32:12Z","timestamp":1678915932616},"reference-count":0,"publisher":"Canadian Science Publishing","issue":"5","license":[{"start":{"date-parts":[[1976,5,1]],"date-time":"1976-05-01T00:00:00Z","timestamp":199756800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/www.nrcresearchpress.com\/page\/about\/CorporateTextAndDataMining"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Can. J. Biochem."],"published-print":{"date-parts":[[1976,5,1]]},"abstract":"<jats:p> A procedure for the quantitative measurement of the O<jats:sup>2\u2032<\/jats:sup>-methylnucleoside constituents of RNA has recently been developed in this laboratory (Gray, M. W. Can. J. Biochem. 53,735\u2013746 (1975)). This assay method is based on the resistance of O<jats:sup>2\u2032<\/jats:sup>-methylnucleoside 5\u2032-phosphates (pNm) (generated by phosphodiesterase hydrolysis of RNA) to subsequent dephosphorylation by venom 5\u2032-nucleotidase (EC 3.1.3.5). In the present investigation, two base-modified 5\u2032-nucleotides, each displaying an unusual resistance to 5\u2032-nucleotidase, have been identified. These compounds have been characterized by a variety of techniques as N<jats:sup>2<\/jats:sup>,N<jats:sup>2<\/jats:sup>-dimethylguanosine 5\u2032-phosphate [Formula: see text] and 3-(3-amino-3-carboxypropyl)uridine 5\u2032-phosphate (p4abu<jats:sup>3<\/jats:sup>U). Because of their resistance to 5\u2032-nucleotidase, [Formula: see text] and p4abu<jats:sup>3<\/jats:sup>U are isolated along with the pNm in the mononucleotide fraction of venom hydrolysates of transfer RNA.Under hydrolysis conditions, the stability of p4abu<jats:sup>3<\/jats:sup>U is comparable to that of a pNm, allowing quantitative assay of the nucleotide. The proportion (mean \u00b1 SD) of p4abu<jats:sup>3<\/jats:sup>U in venom hydrolysates of wheat embryo and Escherichia coli tRNA has been determined to be 0.35\u2002\u00b1\u20020.03 (n\u2002=\u20025) and 0.14\u2002\u00b1\u20020.02 (n\u2002=\u20024) mol%, respectively. The absence of p4abu<jats:sup>3<\/jats:sup>U in venom hydrolysates of yeast tRNA implies the absence of the corresponding nucleoside in yeast tRNA, in agreement with existing data. The variable recovery of [Formula: see text] from venom hydrolysates of wheat embryo and yeast tRNA indicates that under hydrolysis conditions, this base-modified nucleotide is only partially resistant to 5\u2032-nucleotidase. The complete absence of[Formula: see text] in venom hydrolysates of E. coli tRNA is consistent with the known absence of N<jats:sup>2<\/jats:sup>,N<jats:sup>2<\/jats:sup>-dimethylguanosine in this RNA. These observations demonstrate that resistance to 5\u2032-nucleotidase is a necessary but not sufficient criterion for concluding that a 5\u2032-nucleotide is O<jats:sup>2\u2032<\/jats:sup>-methylated.When applied to wheat embryo ribosomal RNA, the analytical methods described in this report failed to reveal any compound having the distinctive charge properties of p4abu<jats:sup>3<\/jats:sup>U. It therefore appears that 1-methyl-3-(3-amino-3-carboxypropyl)pseudouridine, recently characterized as a constituent of the 18 S rRNA of Chinese hamster cells (Saponara, A. G. &amp;Enger, M. D. Biochim. Biophys. Acta 349,61\u201377 (1974)), may not be present in wheat embryo ribosomal RNA. <\/jats:p>","DOI":"10.1139\/o76-060","type":"journal-article","created":{"date-parts":[[2009,12,18]],"date-time":"2009-12-18T21:17:10Z","timestamp":1261171030000},"page":"413-422","source":"Crossref","is-referenced-by-count":1,"title":["Modified 5\u2032-nucleotides resistant to 5\u2032-nucleotidase: isolation of 3-(3-amino-3-carboxypropyl)uridine 5\u2032-phosphate and <i>N<\/i><sup>2<\/sup>,<i>N<\/i><sup>2<\/sup>-dimethylguanosine 5\u2032-phosphate from snake venom hydrolysates of transfer RNA"],"prefix":"10.1139","volume":"54","author":[{"given":"M. W.","family":"Gray","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"155","container-title":["Canadian Journal of Biochemistry"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/www.nrcresearchpress.com\/doi\/pdf\/10.1139\/o76-060","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2020,1,5]],"date-time":"2020-01-05T22:52:46Z","timestamp":1578264766000},"score":1,"resource":{"primary":{"URL":"http:\/\/www.nrcresearchpress.com\/doi\/10.1139\/o76-060"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1976,5,1]]},"references-count":0,"journal-issue":{"issue":"5","published-print":{"date-parts":[[1976,5,1]]}},"alternative-id":["10.1139\/o76-060"],"URL":"https:\/\/doi.org\/10.1139\/o76-060","relation":{},"ISSN":["0008-4018"],"issn-type":[{"value":"0008-4018","type":"print"}],"subject":[],"published":{"date-parts":[[1976,5,1]]}}}