Expression of a chitinase gene and lysis of the host cell wall during Chlorella virus CVK2 infection.
A chitinase gene (vChti-1) encoded by the Chlorella virus CVK2 was cloned and characterized. The vChti-1 open reading frame consisted of 2508 bp corresponding to 836 amino acid residues. The predicted amino acid sequence contained two sets of a family 18 catalytic domain that is responsible for chitinase activity. Northern blot analysis revealed that the vChti-1 gene was expressed in virus-infected Chlorella cells late in infection, when a single transcript of about 2.5 kb appeared at 120 min postinfection. This result was confirmed by Western blotting with a specific anti-vChti-1 protein antibody; a protein of about 94 kDa was detected specifically beginning at 240 min postinfection and was present until cell lysis. The protein was not incorporated into viral particles but remained in the medium after cell lysis. The vChti-1 protein produced in virus-infected cells showed chitinase activity on zymogram assays.
Graduate School of ADSM, Hiroshima University, 1-4-1 Kagamiyama, 739-8527, Japan.
S
S
Hiramatsu
M
M
Ishihara
M
M
Fujie
S
S
Usami
T
T
Yamada
D000595:Amino Acid Sequence; D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D020134:Catalytic Domain; D002473:Cell Wall / Q000378:metabolism*; D002688:Chitinases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D002708:Chlorella / Q000166:cytology / Q000378:metabolism / Q000821:virology*; D003001:Cloning, Molecular; D017124:Conserved Sequence / Q000235:genetics; D005260:Female; D015967:Gene Expression Regulation, Viral; D005814:Genes, Viral / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016366:Open Reading Frames / Q000235:genetics; D017862:Phycodnaviridae / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D013997:Time Factors
D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D002688:Chitinases