The Multigene Family of the Tobacco Hornworm V-ATPase: Novel Subunits A, C, D, H, and Putative Isoforms

Biochim Biophys Acta. 2000 Aug 25;1467(2):369-79. doi: 10.1016/s0005-2736(00)00233-9.

Abstract

The plasma membrane V-ATPase from Manduca sexta (Lepidoptera, Sphingidae) larval midgut is composed of at least 12 subunits, eight of which have already been identified molecularly [Wieczorek et al., J. Bioenerg. Biomembr. 31 (1999) 67-74]. Here we report primary sequences of subunits C, D, H and a, which previously had not been identified in insects. Expression of recombinant proteins, immunostaining and protein sequencing demonstrated that the corresponding proteins are subunits of the Manduca V-ATPase. Genomic Southern blot analysis indicated the existence of multiple genes encoding subunits G, a, c, d and e. Moreover, multiple transcripts were detected in Northern blots from midgut poly(A) RNA for subunits B, G, c and d. Thus, these polypeptides appear to exist as multiple isoforms that could be expressed either in different tissues or at distinct locations within a cell. By contrast subunits A, C, D, E, F and H appear to be encoded by single transcripts and therefore should be present in any Manduca V-ATPase, independent of its subcellular or cell specific origin.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cloning, Molecular
  • DNA Primers / genetics
  • DNA, Complementary / genetics
  • DNA, Complementary / isolation & purification
  • Gene Expression
  • Genes, Insect*
  • Isoenzymes / chemistry
  • Isoenzymes / genetics
  • Manduca / enzymology*
  • Manduca / genetics*
  • Molecular Sequence Data
  • Multigene Family*
  • Protein Structure, Quaternary
  • Proton-Translocating ATPases / chemistry
  • Proton-Translocating ATPases / genetics*
  • Sequence Homology, Amino Acid
  • Vacuolar Proton-Translocating ATPases*

Substances

  • DNA Primers
  • DNA, Complementary
  • Isoenzymes
  • Vacuolar Proton-Translocating ATPases
  • Proton-Translocating ATPases

Associated data

  • GENBANK/AJ249388
  • GENBANK/AJ249389
  • GENBANK/AJ249390
  • GENBANK/AJ251992