Isolation, characterization, and transcriptional analysis of the chitinase chi2 Gene (DQ011663) from the biocontrol fungus Metarhizium anisopliae var. anisopliae.
Metarhizium anisopliae infects arthropods via a combination of specialized structures and cuticle degradation. Hydrolytic enzymes are accepted as key factors for the host penetration step and include chitinases. The characterization of the chi2 chitinase gene from M. anisopliae var. anisopliae is reported. The chi2 gene is interrupted by two short introns and is 1,542-bp long, coding a predicted protein of 419 amino acids with a stretch of 19 amino acid residues displaying characteristics of signal peptide. The predicted chitinase molecular mass is 44 kDa with a mature protein of 42 kDa and a theoretical pI of 4.8. The comparison of the CHI2 predicted protein to fungal orthologues revealed similarity to the glycohydrolase family 18 and a phylogenetic analysis was conducted. The chi2 gene is up-regulated by chitin as a carbon source and in conditions of fungus autolysis, and is down-regulated by glucose. This regulation is consistent with the presence of putative CreA/Crel/Crr1 carbon catabolic repressor binding domains on the regulatory sequence.
Baratto
César Milton
CM
Centro de Biotecnologia, Programa de Pós-graduação em Biologia Celular e Molecular Universidade Federal do Rio Grande do Sul, P. O. Box 15005, 91501-970, Porto Alegre, RS, Brazil.
Dutra
Valéria
V
Boldo
Juliano Tomazzoni
JT
Leiria
Leonardo Barbosa
LB
Vainstein
Marilene Henning
MH
Schrank
Augusto
A
D001483:Base Sequence; D002688:Chitinases / Q000235:genetics*; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D005656:Fungal Proteins / Q000235:genetics; D006999:Hypocreales / Q000201:enzymology / Q000235:genetics*; D008969:Molecular Sequence Data; D010572:Pest Control, Biological; D010802:Phylogeny; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA / Q000379:methods; D024363:Transcription Initiation Site
D004271:DNA, Fungal; D005656:Fungal Proteins; D012333:RNA, Messenger; D002688:Chitinases