title: Mutational analysis of splicing activities of ribonucleotide reductase α subunit protein from lytic bacteriophage P1201.
abstract: A CP1201 RIR1 intein is found in the ribonucleotide reductase alpha subunit (RNR α subunit) protein of lytic bacteriophage P1201 from Corynebacterium glutamicum NCHU 87078. This intein can be over-expressed and spliced in Escherichia coli NovaBlue cells. Mutations of C539, the N-terminal residue of the C-extein in the CP1201 RIR1 protein, led to the changes of pattern and level of protein-splicing activities. A G392S variant was found to be a temperature-sensitive protein with complete splicing activity at 17 and 28°C but not at 37°C or higher. We also found that the cleavage at the CP1201 RIR1 intein C-terminus of the double mutant G392S/C539G was blocked, but other cleavage activities could be efficiently performed at 17°C. G392S/C539G variant possessed the properties of low-temperature-induced cleavage at the intein N-terminus.
authors.affiliation: Institute of Molecular Biology, National Chung Hsing University, Taichung 402, Taiwan.
authors.forename: Shu-Chen
authors.initials: SC
authors.lastname: Kan
authors.forename: Liang-Kun
authors.initials: LK
authors.lastname: Yu
authors.forename: Jiau-Hua
authors.initials: JH
authors.lastname: Chen
authors.forename: Hui-Yu
authors.initials: HY
authors.lastname: Hu
authors.forename: Wen-Hwei
authors.initials: WH
authors.lastname: Hsu
mesh_terms: D020816:Amino Acid Motifs; D001435:Bacteriophages / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D048230:Corynebacterium glutamicum / Q000821:virology; D004795:Enzyme Stability; D047668:Inteins; D008969:Molecular Sequence Data; D009154:Mutation; D021122:Protein Subunits / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012326:RNA Splicing; D012264:Ribonucleotide Reductases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
chemical_list: D021122:Protein Subunits; D014764:Viral Proteins; D012264:Ribonucleotide Reductases