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Arch Virol ; 149(2): 241-59, 2004 Feb.
Article in English | MEDLINE | ID: mdl-14745593

ABSTRACT

Late during the bacteriophage Mu lytic cycle, Mu DNA must be matured and packaged from its dispersed integration sites in the host DNA in order to produce progeny virions. Whereas control of late gene transcription in Mu is becoming well understood, less is known about the phage morphogenetic process. To investigate the latter, we cloned and sequenced a approximately 4.3-kb region of the phage DNA beginning just upstream of the leftmost late promoter Plys. Previous mapping of amber mutations had located the lysis (lys) and proposed DNA maturation genes D and E in this region. When the DNA sequence was analyzed, seven potential open reading frames were found. DNA sequence analysis of amber mutations in genes D and E identified the sixth and seventh open reading frames as D and E, respectively. Cloning and expression of this region enabled production of cell-free protein extracts that specifically recognize the phage-encoded packaging sequence (pac), a characteristic exhibited by phage maturation enzymes. In addition, the E protein was found to share homology with the large subunit of many phage DNA maturation enzymes. These results support the hypothesis that D and E encode subunits of the Mu DNA maturation enzyme.


Subject(s)
Bacteriophage mu/genetics , DNA Transposable Elements/genetics , Gene Order/genetics , Genes, Viral/genetics , Amino Acid Motifs , Amino Acid Sequence , Bacteriophage mu/enzymology , DNA-Binding Proteins/chemistry , DNA-Binding Proteins/genetics , Electrophoretic Mobility Shift Assay , Endodeoxyribonucleases/chemistry , Endodeoxyribonucleases/genetics , Escherichia coli/virology , Hydrophobic and Hydrophilic Interactions , Molecular Sequence Data , Mutation/genetics , Open Reading Frames/genetics , Promoter Regions, Genetic/genetics , Sequence Alignment
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