Biomaterial Database

Identification of the duplicated genes for S-adenosyl-l-methionine synthetase (metK1-sp and metK2-sp) in Streptomyces peucetius var. caesius ATCC 27952. 2010

T-J Oh, and N P Niraula, and K Liou, and J K Sohng

Institute of Biomolecule Reconstruction (iBR), Department of Pharmaceutical Engineering, Sun Moon University, Tangjeong-myeon, Asansi, Chungnam, Republic of Korea.

OBJECTIVE To characterize the function of both metK1-sp (sp1190) and metK2-sp (sp1566) in vitro and in vivo, and to study the regulation of doxorubicin production by overexpressing the metK. RESULTS We cloned two orfs into pET32a(+) respectively, and the formation of S-Adenosyl-l-methionine was clearly observed in the in vitro enzyme assays as functional MetKs. Reverse transcriptase polymerase chain reaction (PCR) analysis indicated that the transcripts for the metK1-sp were repressed as Streptomyces cells entered the decline phase, whereas that of the metK2-sp was induced, suggesting that these MetK proteins may be important for the growth and the regulation of secondary metabolites during the stationary growth phase, whether considered together or separately. Furthermore, we found that the introduction of high-copy-number plasmids containing the metK1-sp and metK2-sp resulted in 2.1- and 1.4-fold greater levels of doxorubicin production than the control transformants containing only the vector, respectively. We also attempted to disrupt the metK-sp and found that doxorubicin production from the metK1-sp-deleted mutant (Streptomyces peucetius/pNN1) was reduced when compared to the parent strain (S. peucetius var. caesius ATCC 27952). CONCLUSIONS The results of this study indicated that two metK are differentially expressed during cell growth, and that the expressions of the two metK genes are differentially regulated under the same conditions. CONCLUSIONS Streptomyces peucetius var. caesius contains two genes, metK1-sp and metK2-sp, which encode functional S-adenosyl-l-methionine synthetase (MetK). The degree of homology (90% identity) found between the two genes shows that metK1-sp and metK2-sp are duplicated genes. Although there is currently no evidence for the relationship of the duplicated metK genes involved in the regulation of doxorubicin production, metK1-sp and metK2-sp may play a role in controlling the stimulation of antibiotic production during secondary metabolism.

UI	MeSH Term	Description	Entries
D008716	Methionine Adenosyltransferase	An enzyme that catalyzes the synthesis of S-adenosylmethionine from methionine and ATP. EC 2.5.1.6.	S-Adenosylmethionine Synthetase,ATP-Methionine S-Adenosyltransferase,ATP Methionine S Adenosyltransferase,Adenosyltransferase, Methionine,S Adenosylmethionine Synthetase,S-Adenosyltransferase, ATP-Methionine,Synthetase, S-Adenosylmethionine
D008969	Molecular Sequence Data	Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories.	Sequence Data, Molecular,Molecular Sequencing Data,Data, Molecular Sequence,Data, Molecular Sequencing,Sequencing Data, Molecular
D004317	Doxorubicin	Antineoplastic antibiotic obtained from Streptomyces peucetius. It is a hydroxy derivative of DAUNORUBICIN.	Adriamycin,Adriablastin,Adriablastine,Adriblastin,Adriblastina,Adriblastine,Adrimedac,DOXO-cell,Doxolem,Doxorubicin Hexal,Doxorubicin Hydrochloride,Doxorubicin NC,Doxorubicina Ferrer Farm,Doxorubicina Funk,Doxorubicina Tedec,Doxorubicine Baxter,Doxotec,Farmiblastina,Myocet,Onkodox,Ribodoxo,Rubex,Urokit Doxo-cell,DOXO cell,Hydrochloride, Doxorubicin,Urokit Doxo cell
D000595	Amino Acid Sequence	The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION.	Protein Structure, Primary,Amino Acid Sequences,Sequence, Amino Acid,Sequences, Amino Acid,Primary Protein Structure,Primary Protein Structures,Protein Structures, Primary,Structure, Primary Protein,Structures, Primary Protein
D013302	Streptomyces	A genus of bacteria that form a nonfragmented aerial mycelium. Many species have been identified with some being pathogenic. This genus is responsible for producing a majority of the ANTI-BACTERIAL AGENTS of practical value.
D017386	Sequence Homology, Amino Acid	The degree of similarity between sequences of amino acids. This information is useful for the analyzing genetic relatedness of proteins and species.	Homologous Sequences, Amino Acid,Amino Acid Sequence Homology,Homologs, Amino Acid Sequence,Homologs, Protein Sequence,Homology, Protein Sequence,Protein Sequence Homologs,Protein Sequence Homology,Sequence Homology, Protein,Homolog, Protein Sequence,Homologies, Protein Sequence,Protein Sequence Homolog,Protein Sequence Homologies,Sequence Homolog, Protein,Sequence Homologies, Protein,Sequence Homologs, Protein
D058977	Molecular Sequence Annotation	The addition of descriptive information about the function or structure of a molecular sequence to its MOLECULAR SEQUENCE DATA record.	Gene Annotation,Protein Annotation,Annotation, Gene,Annotation, Molecular Sequence,Annotation, Protein,Annotations, Gene,Annotations, Molecular Sequence,Annotations, Protein,Gene Annotations,Molecular Sequence Annotations,Protein Annotations,Sequence Annotation, Molecular,Sequence Annotations, Molecular
D020131	Genes, Duplicate	Two identical genes showing the same phenotypic action but localized in different regions of a chromosome or on different chromosomes. (From Rieger et al., Glossary of Genetics: Classical and Molecular, 5th ed)	Duplicate Genes,Duplicate Gene,Gene, Duplicate