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Vol. 18, Art. 13 (pp. 200-208)    |    2017       

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Small subunit of restriction endonuclease BspD6I is inactive in the presence of catalytic-deficient large subunit

Yunusova A.K.1, Artyukh R.I.1, Perevyazova T.A.1, Abrosimova L.A.3,
Kachalova G.S.2, Zheleznaya L.A.1

1 Institute of Theoretical and Experimental Biophysics of Russian Academy of Science

2 Federal Research Centre "Fundamentals of Biotechnology" of the Russian Academy of Sciences

3 Chemistry Department and A.N. Belozersky Institute of Physico-Chemical Biology

Brief summary

The large subunit of heterodimeric restriction endonuclease BspD6I on its own binds to DNA and specifically nicks the top strand of its recognition sequence. The small subunit of BspD6I is inactive on its own but in the presence of the large subunit cleaves the top strand of the recognition sequence. To elucidate what the large subunit properties are important for displaying small subunit activity two catalytic-deficient large subunits (D456A и E418A) we received. The results show that small subunit is active only in the presence of catalytic active large subunit. It is proposed that catalytic active large subunit binding DNA takes on special orientation and the small becomes able to hydrolyze the bottom strand of DNA.


Key words

heterodimeric restriction endonuclease, nicking endonulease, nickase, mutagenesis

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