Cloning and characterization of a ribonuclease L inhibitor from the silkworm, Bombyx mori.
The ribonuclease L (RNase L) pathway plays an important role in the response of cells to double-stranded RNA (dsRNA) during the events such as virus infection. Ribonuclease L inhibitor (RLI) belonging to the ABC transporter family is known as a regulator of the RNase L pathway. The homologs of RLI were reported in many organisms including the fruit fly and mosquito, but their functions in insects and arthropods have not been elucidated to date. In the present study, we cloned a cDNA of a silkworm RLI homolog, termed BmRLI, and its nucleotide sequence was determined. RT-PCR analysis revealed that the expression of BmRLI mRNA was marked in the testis, ovary and fat body. From the cDNA, recombinant protein with an apparent molecular mass of 69 kDa was expressed in Escherichia coli and cultured insect cells. Although no obvious effect of up-regulation of the BmRLI expression on RNAi was observed, its down-regulation slightly reduced RNAi efficiency.
Maeda
Takuji
T
Laboratory of Silkworm Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, Fukuoka, Japan.
Lee
Jae Man
JM
Miyagawa
Yoshitaka
Y
Koga
Katsumi
K
Kawaguchi
Yutaka
Y
Kusakabe
Takahiro
T
D018528:ATP-Binding Cassette Transporters / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D012831:Bombyx / Q000235:genetics*; D018833:Chaperonins / Q000235:genetics*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004722:Endoribonucleases / Q000037:antagonists & inhibitors*; D005216:Fat Body / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D017344:Genes, Insect; D019476:Insect Proteins / Q000235:genetics*; D008297:Male; D008969:Molecular Sequence Data; D010053:Ovary / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013737:Testis / Q000378:metabolism; D014018:Tissue Distribution
D018528:ATP-Binding Cassette Transporters; D018076:DNA, Complementary; D019476:Insect Proteins; D012333:RNA, Messenger; D004722:Endoribonucleases; C031956:2-5A-dependent ribonuclease; D018833:Chaperonins