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5-重氮尿嘧啶-2-C掺入丝状大肠杆菌核糖核酸中标记残基的表征

Characterization of Labeled Residues from 5-Diazouracil-2-C Incorporated into Ribonucleic Acids of Filamentous Escherichia coli.

作者信息

Fister G, Previc E, Richardson S

机构信息

Department of Microbiology, University of Florida, Gainesville, Florida 32601.

出版信息

J Bacteriol. 1970 Dec;104(3):1213-22. doi: 10.1128/jb.104.3.1213-1222.1970.

Abstract

E. coli B, filamented with 5-diazouracil (DZU)-2-(14)C, yielded ribonucleic acid (RNA)-(DZU-2-(14)C) which was converted by pancreatic ribonuclease to (14)C-mono-and oligo-nucleotides. The mixed (14)C-mononucleotides isolated by diethylaminoethyl-cellulose fractionation were identified as cytidylic, uridylic, and hydroxyuridylic acids, by using a combination of paper chromatography and treatment with alkaline phosphatase and cytidine deaminase. Rifampin blocked incorporation of DZU-2-(14)C under conditions which inhibit RNA synthesis. Division inhibition by DZU-2-(14)C and the incorporation into Escherichia coli B were retarded by uracil but not by other RNA bases. In a pyrimidine-requiring E. coli, DZU substituted for uracil or cytosine to an extent limited by toxic effects. Cytosine and uracil retarded these effects and retarded the incorporation of DZU-2-(14)C into the pyrimidineless strain. A small proportion of DZU-2-(14)C was converted by the latter strain into hydroxyuridylic acid, but the bulk of the incorporated label was in cytidylic and uridylic acid, as in the wild strain.

摘要

用5-重氮尿嘧啶(DZU)-2-(14)C使大肠杆菌B丝状化,产生核糖核酸(RNA)-(DZU-2-(14)C),其经胰核糖核酸酶转化为(14)C-单核苷酸和寡核苷酸。通过二乙氨基乙基纤维素分级分离得到的混合(14)C-单核苷酸,通过纸色谱法、碱性磷酸酶处理和胞苷脱氨酶处理相结合的方法,被鉴定为胞苷酸、尿苷酸和羟基尿苷酸。在抑制RNA合成的条件下,利福平阻断了DZU-2-(14)C的掺入。DZU-2-(14)C对细菌分裂的抑制作用以及其掺入大肠杆菌B的过程,受到尿嘧啶的阻碍,但不受其他RNA碱基的影响。在一株需要嘧啶的大肠杆菌中,DZU替代尿嘧啶或胞嘧啶的程度受到毒性作用的限制。胞嘧啶和尿嘧啶会延缓这些作用,并延缓DZU-2-(14)C掺入无嘧啶菌株。后一株菌株将一小部分DZU-2-(14)C转化为羟基尿苷酸,但与野生菌株一样,掺入的大部分标记物存在于胞苷酸和尿苷酸中。

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