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J. Biochem, 1984, Vol. 96, No. 5 1501-1509
© 1984 Japanese Biochemical Society

research-article

Utilizations of Various Uridine 5' -Triphosphate Analogues by DNA-Dependent RNA Polymerases I and II Purified from Liver Nuclei of the Cherry Salmon (Oncorhynchus masou)1

Chikao NAKAYAMA and Mineo SANEYOSHI

Faculty of Pharmaceutical Sciences, Hokkaido University, Kita-ku Sapporo, Hokkaido 060

Various 5-substituted UTPs (methyl, ethyl, n-propyl, n-butyl, fluoro, chloro, bromo, and iodo) and sulfur-containing UTP analogues (4-thio-, 2-thio-, 5-methyl-2-thio-, and 5-methyl-4-thio-) were synthesized chemically and their utilization by DNA-dependent-RNA polymerases I and II of the cherrry salmon (Oncorhynchus masou) were studied in substitution experiments under the condition of limited RNA synthesis in vitro. RNA polymerase I utilized the 5-methyl-, chloro, bromo, and iodo derivatives of UTP more efficiently than unmodified UTP, but RNA polymerase II utilized UTP most efficiently. 5-Methyl-4-thiouridine 5'-triphosphate (4-thio TTP) was utilized more efficiently than UTP by RNA polymerase I. On the other hand, it was found that 4-thio TTP was a selective substrate for RNA polymerase I and that its incorporation by RNA polymerase II was very slow. Thus recognition of UTP analogues as substrates by RNA polymerase I and II was different. These observation were attributed from kinetic analyses to differences in catalytic activity(Vmax).

1 This work was partly supported by a Grant-in-Aid of Cancer Research from the Ministry of Education, Science and Culture of Japan to M. S.


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