A DnaJ-like homolog from Cryphonectria parasitica is not responsive to hypoviral infection but is important for fungal growth in both wild-type and hypovirulent strains.
A DnaJ-like gene, Cpdj1, a molecular chaperone and regulator of Hsp70 in Cryphonectria parasitica, was characterized. The protein product of Cpdj1 gene consists of 379 amino acids with a predicted molecular mass of 40.6 kDa and a pI of 7.79. The deduced protein sequence revealed preservation of the conserved hall-mark J-region and exhibited high homolo y to all known DnaJ-like proteins. Disruption of the Cpdj1 gene resulted in slow growth and produced colonies characterized by retarded growth and deep orange color. Accordingly, reduced virulence of the Cpdj1-null mutant was observed. This reduced growth rate was magnified when the Cpdj1-null mutant was cultured under heat-stress conditions. Reduced conidiation was also observed in the Cpdj1-null mutant, indicating that Cpdj1 gene, although not essential for cell viability, is required for appropriate cellular processes including growth and sporulation. Northern analysis showed that Cpdj1 was constitutively expressed, and when the culture was subject to high temperature, a strong induction of the transcript was observed. No significant difference in the expression and induction pattern of Cpdj1 was observed between virus-free EP155/2 and virus-infected hypovirulent UEP1 strains. However, further severe defects in mycelia growth and conidiation were observed in the hypovirus-infected Cpdj1-null mutant suggesting that the presence of Cpdj1 is required for mycelia growth and sporulation of the hypovirus-infected strain.
Lim
Jung-Gu
JG
Institute for Molecular Biology and Genetics, Center for Fungal Pathogenesis, Chonbuk National University, Jeonju, 561-756, Korea.
Lee
Joong-Gi
JG
Kim
Jung-Mi
JM
Park
Jin-Ah
JA
Park
Seung-Moon
SM
Yang
Moon-Sik
MS
Kim
Dae-Hyuk
DH
D000595:Amino Acid Sequence; D048708:Cell Growth Processes / Q000235:genetics; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D005658:Fungi / Q000254:growth & development / Q000502:physiology*; D015966:Gene Expression Regulation, Fungal; D005816:Genetic Complementation Test; D050956:HSP40 Heat-Shock Proteins / Q000235:genetics* / Q000378:metabolism; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics; D018869:Heat-Shock Response / Q000235:genetics; D006358:Hot Temperature / Q000009:adverse effects; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D025282:Mycelium / Q000235:genetics / Q000254:growth & development; D017386:Sequence Homology, Amino Acid; D013172:Spores, Fungal / Q000235:genetics; D014774:Virulence / Q000235:genetics; D014777:Virus Diseases / Q000235:genetics / Q000378:metabolism*
D005656:Fungal Proteins; D050956:HSP40 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins