Molecular cloning and expression of a hexamerin cDNA from the honey bee, Apis mellifera.
A cDNA encoding a hexamerin subunit of the Africanized honey bee (Apis mellifera) was isolated and completely sequenced. In the deduced translation product we identified the N-terminal sequence typical of the honey bee HEX 70b hexamerin. The genomic sequence consists of seven exons flanked by GT/AT exon/intron splicing sites, which encode a 683 amino acid polypeptide with an estimated molecular mass of 79.5 kDa, and pI value of 6.72. Semi-quantitative RT-PCR revealed high levels of Hex 70b message in larval stages, followed by an abrupt decrease during prepupal-pupal transition. This coincides with decaying titers of juvenile hormone (JH) and ecdysteroids that is the signal for the metamorphic molt. To verify whether the high Hex 70b expression is dependent on high hormone levels, we treated 5th instar larvae with JH or 20-hydroxyecdysone (20E). In treated larvae, Hex 70b expression was maintained at high levels for a prolonged period of time than in the respective controls, thus indicating a positive hormone regulation at the transcriptional level. Experiments designed to verify the influence of the diet on Hex 70b expression showed similar transcript amounts in adult workers fed on a protein-enriched diet or fed exclusively on sugar. However, sugar-fed workers responded to the lack of dietary proteins by diminishing significantly the amount of HEX 70b subunits in hemolymph. Apparently, they use HEX 70b to compensate the lack of dietary proteins.
Cunha
Adriana D
AD
Departamento de Biologia, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, Av. Bandeirantes 3900, Ribeirão Preto 14040-901, SP, Brazil.
Nascimento
Adriana M
AM
Guidugli
Karina R
KR
Simões
Zilá L P
ZL
Bitondi
Márcia M G
MM
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001516:Bees / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D004441:Ecdysterone / Q000494:pharmacology; D015870:Gene Expression; D006458:Hemolymph / Q000737:chemistry; D019476:Insect Proteins / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D010802:Phylogeny; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D012717:Sesquiterpenes / Q000494:pharmacology
D018076:DNA, Complementary; D019476:Insect Proteins; D012717:Sesquiterpenes; C041137:storage proteins, Insecta; D004441:Ecdysterone; C036585:juvenile hormone III