Ribonucleotide Reductase Activity of Schizosaccbarornyces pombe Is Inhibited by Escherichia coli RecA Antibody

Schizosaccbarornyces pombe에서 Escherichia coli RecA 항체에 의한 Ribonucleotide Reductase 효소활성 저해

We have previoosly demonstrated that the RecA-like protein of Schizosaccharomyces pombe (S. pombe) is immunologically related to Escherichia coil (E. coil) RecA protein and that the cellular level of the protein is significantly increased by inhibitors of nucleotide pool-forming enzymes such as hydroxyurea (HU) and methotrexate (MTX) (lee and Park, 1994; lee et al., 1994). In this study, we report that the ribonudeotide redudase activity of S. pombe is inhibited by E. coil RecA antibody, as determined by thin layer chromatography using [5-$^3$H]CDP as a substrate. The relative activity of ribonucleotide reductase was dramatically inhibited by 100 mM of flu (26.4% reduction) in in vitro assay, compared to that of non-treated control. The ribonucleotide reductase activity was also inhibited by immunoprecipitation with E. coil RecA antibody (43.3% reduction). These results indicate that the strudure of S. pombe ribonucleotide reductase is in part similar to that of E. coil RecA protein.

분열형 호모인 Schizosaccbarornyces pombe에서 RecA 유사 단백질은 hydroxyurea (HU)나 methotrexate (MTX)와 같은 nucleotide pool 형성에 관여하는 효소의 저해제에 의해 유도발현된다. 본 연구에서는 대장균의 RecA 단백질과 S. pombe의 ribonucleotide reductase가 구조적으로 유사한지를 RecA 항체를 이용한 면역침전법과 [5-$^3$H]CDP를 기질로하는 thin layerchromatography방법을 통하여 알아보고자 하였다. S. pombe의 rinonucleotide reductase활성은 100mM HU에 의해 대조군에 비해 26.3% 저해되었으며, RecA 항체를 이용한 면역침전에 의해서도 43.3% 저해되었다. 이와 같은 결과는 S. pombe의 rinonucleotide reductase가 대장균의 RecA 단백질과 구조적으로 유사함을 시사하는 것이다.