The Corynebacterium glutamicum cglIM gene encoding a 5-cytosine methyltransferase enzyme confers a specific DNA methylation pattern in an McrBC-deficient Escherichia coli strain |
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| Institution: | 1. PhD Course in Oncology and Experimental Surgery, University of Palermo, Palermo, Italy;2. “P. Giaccone” University Hospital, School of Medicine, University of Palermo, Palermo, Italy;3. Department of Biologic, Chemical, and Pharmaceutical Sciences and Technologies, University of Palermo, Palermo, Italy;4. “A. Mirri” Zooprophylactic Institute of Sicily, Palermo, Italy;5. Dichirons Department, University of Palermo, Palermo, Italy;1. Moscow Institute of Physics and Technology (State University), Moscow Region, Dolgoprudny, 141701, Russia;2. V.L.Talrose Institute for Energy Problems of Chemical Physics, Russian Academy of Sciences, Moscow, 119334, Russia;3. Institute of Biomedical Chemistry, Moscow, 119121, Russia;1. School of Chemical Sciences, Central University of Gujarat, Gandhinagar 382030, Gujarat, India;2. Department of Biochemistry, National JALMA Institute for Leprosy and Other Mycobacterial Diseases (ICMR), Taj Ganj, Agra 282004, India;3. Centre for Applied Chemistry, Central University of Gujarat, Gandhinagar 382030, Gujarat, India;1. Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, 159 Long Pan Road, Nanjing 210037, China;2. College of Chemical Engineering, Nanjing Forestry University, 159 Long Pan Road, Nanjing 210037, China;3. Jiangsu Key Lab for the Chemistry & Utilization of Agricultural and Forest Biomass, 159 Long Pan Road, Nanjing 210037, China;4. Jiangsu Kanion Pharmaceutical Co., Ltd., 58 Haichang South Road, Lianyungang 222001, Jiangsu Province, China |
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| Abstract: | The cglIM gene of the coryneform soil bacterium Corynebacterium glutamicum ATCC 13032 has been cloned and characterized. The coding region comprises 1092 nucleotides and specifies a protein of 363 amino acid residues with a deduced Mr of 40 700. The amino acid sequence showed striking similarities to methyltransferase enzymes generating 5-methylcytosine residues, especially to M·NgoVII from Neisseria gonorrhoeae recognizing the sequence GCSGC. The cglIM gene is organized in an unusual operon which contains, in addition, two genes encoding stress-sensitive restriction enzymes. Using PCR techniques the entire gene including the promoter region was amplified from the wild-type chromosome and cloned in Escherichia coli. Expression of the cglIM gene in E. coli under the control of its own promoter conferred the C. glutamicum-specific methylation pattern to co-resident shuttle plasmids and led to a 260-fold increase in the transformation rate of C. glutamicum. In addition, the methylation pattern produced by this methyltransferase enzyme is responsible for the sensitivity of DNA from C. glutamicum to the modified cytosine restriction (Mcr) system of E. coli. |
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