The Tsp45I restriction-modification system is plasmid-borne within its thermophilic host

Gene
J WayneS Y Xu

Abstract

Thermus species YS45 harbors two small cryptic plasmids of 5.8 (pTsp45s) and approximately 12 kb (pTsp45I). Plasmid pTsp45s has been entirely sequenced, revealing three significant ORFs. In addition to a previously reported thermophilic plasmid-encoded replication protein (Rep), pTsp45s contains two genes for the Tsp45I methyltransferase (M.Tsp45I) and restriction endonuclease (Tsp45I). These two converging genes (tsp45IM and tsp45IR) overlap by 4 bp at their stop codons within an XbaI site. M.Tsp45I (413 aa, 47.0 kDa, recognizing 5'-GTSAC-3') is highly homologous to other m6A-methyltransferases, especially M.EcaI (recognizing 5'-GGTNACC-3'). Tsp45I (332 aa, 37.4 kDa, cleaving 5'-/GTSAC-3') is not homologous to M.Tsp45I, or to other restriction endonucleases. Recombinant Tsp45I is stably produced in E. coli, and cleaves DNA at 65 degrees C with the same specificity as the native enzyme. Therefore, the thermophilic Tsp45I restriction-modification system is plasmid-borne within its native host.

References

Feb 1, 1978·Journal of General Microbiology·F HishinumaK Sakaguchi
Jan 1, 1991·Annual Review of Genetics·G G Wilson, N E Murray
Feb 1, 1991·Journal of Bacteriology·T TaoR M Blumenthal
May 25, 1991·Nucleic Acids Research·G G Wilson
Aug 25, 1988·Nucleic Acids Research·N V MurzinaN I Matvienko
Jan 1, 1997·Nucleic Acids Research·R J Roberts, D Macelis

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