Molecular characterization of MbADGF, a novel member of the adenosine deaminase-related growth factor in the cabbage armyworm, Mamestra brassicae: the functional roles in the midgut cell proliferation.
To clarify the functional mechanism of the adenosine deaminase-related growth factor (ADGF) particularly in the regulation of insect development, the cDNA encoding a homologue of ADGF proteins was cloned from the cabbage armyworm, Mamestra brassicae, named MbADGF. The purified MbADGF recombinant protein stimulated cell proliferation in a dose-dependent manner of SES-MaBr-4 and NIAS-MaBr-93 cell lines that were derived from fat bodies and haemocytes of M. brassicae. The adenosine deaminase activity of MbADGF was detected using adenosine and 2'-deoxyadenosine as substrates. Northern analysis demonstrated that during the larval development the level of MbADGF in the midgut increased. In situ hybridization showed that MbADGF mRNA was expressed in midgut goblet cells and in the apical cytoplasm of columnar cells, which suggests that MbADGF protein may execute its adenosine deaminase activity at the apical cytoplasm of columnar cells to convert adenosine into inosine.
Zhang
J
J
Division of Biofunctional Science, Graduate School of Science and Technology, Kobe University, Nada, Kobe, Japan. zhjunmba@hotmail.com
Takeda
M
M
D000241:Adenosine / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects*; D016000:Cluster Analysis; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D003839:Deoxyadenosines / Q000378:metabolism; D004064:Digestive System / Q000166:cytology* / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D017403:In Situ Hybridization; D019476:Insect Proteins / Q000235:genetics*; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics*; D007814:Larva / Q000378:metabolism; D008969:Molecular Sequence Data; D009036:Moths / Q000235:genetics* / Q000378:metabolism; D010802:Phylogeny; D011994:Recombinant Proteins / Q000235:genetics / Q000494:pharmacology*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D018076:DNA, Complementary; D003839:Deoxyadenosines; D019476:Insect Proteins; D036341:Intercellular Signaling Peptides and Proteins; D011994:Recombinant Proteins; D000241:Adenosine; C058118:2'-deoxyadenosine