Characterization and expression of the Hex 110 gene encoding a glutamine-rich hexamerin in the honey bee, Apis mellifera.
An N-terminal amino acid sequence of a previously reported honey bee hexamerin, HEX 110 [Danty et al., Insect Biochem Mol Biol 28:387-397 (1998)], was used as reference to identify the predicted genomic sequence in a public GenBank database. In silico analysis revealed an ORF of 3,033 nucleotides that encompasses eight exons. The conceptual translation product is a glutamine-rich polypeptide with a predicted molecular mass of 112.2 kDa and pI of 6.43, which contains the conserved M and C hemocyanin domains. Semiquantitative and quantitative RT-PCR with specific primers allowed for an analysis of mRNA levels during worker bee development and under different physiological conditions. Concomitantly, the abundance of the respective polypeptide in the hemolymph was examined by SDS-PAGE. Hex 110 transcripts were found in high levels during the larval stages, then decreased gradually during the pupal stage, and increased again in adults. HEX 110 subunits were highly abundant in larval hemolymph, decreased at the spinning-stage, and remained at low levels in pupae and adults. In 5th instar larvae, neither starvation nor supplementation of larval food with royal jelly changed the Hex 110 transcript levels or the amounts of HEX 110 subunit in hemolymph. In adult workers, high levels of Hex 110 mRNA, but not of the respective subunit, were related to ovary activation, and also to the consumption of a pollen-rich diet.
Bitondi
Márcia M G
MM
Departamento de Biologia, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, Brazil. mmgbit@usp.br
Nascimento
Adriana M
AM
Cunha
Adriana D
AD
Guidugli
Karina R
KR
Nunes
Francis M F
FM
Simões
Zilá L P
ZL
D000595:Amino Acid Sequence; D000818:Animals; D001516:Bees / Q000235:genetics* / Q000254:growth & development; D005227:Fatty Acids / Q000378:metabolism; D005260:Female; D018507:Gene Expression Regulation, Developmental; D005973:Glutamine / Q000378:metabolism; D006458:Hemolymph / Q000737:chemistry; D019476:Insect Proteins / Q000737:chemistry* / Q000235:genetics*; D007814:Larva / Q000235:genetics; D008969:Molecular Sequence Data; D010053:Ovary / Q000378:metabolism; D011058:Pollen / Q000378:metabolism; D011679:Pupa / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D000099219:Royal Jelly
D005227:Fatty Acids; D005973:Glutamine; D019476:Insect Proteins; D012333:RNA, Messenger; D000099219:Royal Jelly; C041137:storage proteins, Insecta