Sense and antisense RNA for the membrane associated 40 kDa subunit M40 of the insect V-ATPase.
For the first time a cDNA encoding the membrane associated subunit M40 of an invertebrate V-ATPase has been isolated and sequenced, based on a cDNA library from larval midgut of the tobacco hornworm, Manduca sexta. Immunoblotting with monospecific antibodies raised against the recombinant M40 polypeptide demonstrated that it is a subunit of the insect plasma membrane V-ATPase. Since M40 subunits had been identified only in endosomal V-ATPases till now, this result indicates that they are constitutive members of all, endomembrane and plasma membrane V-ATPases. A phagemid clone representing a polyadenylated antisense transcript was also isolated and sequenced. Using RT-PCR, endogenous antisense RNA was detected in poly(A) RNA isolated from the larval midgut. Since Southern blots indicated a single gene locus, both the antisense RNA as well as the sense mRNA encoding subunit M40 seem to originate from the same gene.
Merzendorfer
H
H
Zoologisches Institut der Universität, München, Germany.
Harvey
W R
WR
Wieczorek
H
H
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D015153:Blotting, Western; D002462:Cell Membrane / Q000201:enzymology; D003001:Cloning, Molecular; D016373:DNA, Antisense / Q000235:genetics; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D018402:Manduca / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D006180:Proton-Translocating ATPases / Q000737:chemistry / Q000235:genetics*; D016372:RNA, Antisense / Q000235:genetics*; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D025262:Vacuolar Proton-Translocating ATPases
D016373:DNA, Antisense; D018076:DNA, Complementary; D016372:RNA, Antisense; D012333:RNA, Messenger; D025262:Vacuolar Proton-Translocating ATPases; D006180:Proton-Translocating ATPases