The structural protein E of the archaeal virus phiCh1: evidence for processing in Natrialba magadii during virus maturation.
phiCh1 is a lysogenic virus for the haloalkalophilic archaeon Natrialba magadii. The virus morphology resembles other members of Myoviridae infecting Halobacterium species. The gene of the major capsid protein E of virus phiCh1 was cloned and the DNA sequence was determined. Gene E was mapped to a 3.2-kbp ClaI fragment, localized to the 5'-end of the phiCh1 genome. The complete nucleotide sequence of this region was determined and the identity of gene E was confirmed by comparing the experimentally determined N-terminal amino acid sequence of the purified protein to the translated DNA sequence of its open reading frame. We present evidence that the gene E product is proteolytically cleaved between Lys(16) and Asn(17) to yield the 305 residue polypeptides found in the mature viral capsid. Processing of the protein itself during virus development was determined by 2D gel electrophoresis using protein E-specific antibodies. Sequence similarity studies revealed an 80% identity to capsid protein Hp32 of phiH, infecting Halobacterium salinarum. RT-PCR analysis as well as Western blot studies revealed gene E as a late gene. Transcripts and proteins could be detected shortly before onset of lysis of the lysogenic strain N. magadii L11.
Klein
R
R
Institute of Microbiology and Genetics, University of Vienna, Dr. Bohr-Gasse 9, Vienna, A-1030, Austria.
Greineder
B
B
Baranyi
U
U
Witte
A
A
D000595:Amino Acid Sequence; D002213:Capsid / Q000235:genetics / Q000378:metabolism*; D036022:Capsid Proteins; D003001:Cloning, Molecular; D004279:DNA, Viral; D004926:Escherichia coli; D020869:Gene Expression Profiling; D006216:Halobacteriaceae / Q000378:metabolism*; D008969:Molecular Sequence Data; D017900:Myoviridae / Q000235:genetics / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D017422:Sequence Analysis, DNA; D014779:Virus Replication
D036022:Capsid Proteins; D004279:DNA, Viral; C415769:protein E, phich1 virus