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Isolation and characterization of Neurospora crassa nucleoside diphosphate kinase NDK-1 by Yasunobu Ogura; Yusuke Yoshida; Kazuya Ichimura; Chito Aoyagi; Naoto Yabe; Kohji Hasunuma is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

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We have previously reported that phosphorylation of a 15‐kDa protein increased after blue‐light irradiation in __Neurospora crassa__. In this study, the 15‐kDa protein was purified using four columns; DEAE–cellulose, Blue–Sepharose, SP–Sepharose and Mono Q. The 15‐kDa protein was shown to be homologous with nucleoside diphosphate kinase by amino acid sequencing and was also shown to possess nucleoside diphosphate kinase activity. A gene encoding __N. crassa__ nucleoside diphosphate kinase, __ndk‐1__, was isolated from the mycelial cDNA and genomic libraries. The deduced amino acid sequence of NDK‐1 was identical to that of the 15‐kDa protein. Northern blot analysis suggested that WC‐1 and WC‐2, the key factors of blue‐light signal transduction in __N. crassa__, did not regulate __NDK‐1__ at the transcriptional level. NDK‐1 also showed rapid autophosphorylation activity and protein kinase activity against myelin basic protein with a __K__~m~ value of 0.36 mm. These results suggest that NDK‐1 acts as a signal transducer by phosphorylating proteins.

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Author

Yasunobu Ogura; Yusuke Yoshida; Kazuya Ichimura; Chito Aoyagi; Naoto Yabe; Kohji Hasunuma

Publisher

John Wiley and Sons; Springer; Wiley (Blackwell Publishing); Springer-Verlag; Blackwell Publishing Inc.; Springer Verlag; Wiley; Springer Science and Business Media LLC (ISSN 1432-1327)

Published

1999

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)