Abstract
The posterior section of Galleria mellonella silk glands contains two abundant mRNAs that are identical except for the non-coding tail, which includes either two (1.1 kb mRNA) or three (1.2 kb mRNA) consensus sequences for polyadenylation sites. The transcripts are 40% homologous in the coding as well as non-coding regions with the mRNA encoding light-chain fibroin (L-fibroin) in Bombyx mori; the deduced translation product shows 43% identity with the Bombyx L-fibroin peptide, with all three cysteines conserved. Amino acid analysis of the N-termini of Galleria silk proteins revealed that L-fibroin (25 kDa) occurs in two isoforms, the shorter one lacking the Ala-Pro dipeptide residue at its N-terminus. The 29 and 30 kDa Galleria silk proteins appear to be homologs of Bombyx silk component P25. The results suggest that evolutionary diversification of Galleria and Bombyx L-fibroins involves alternative polyadenylation and proteolytic processing sites.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Boman HG, Boman IA, Andreu D, Li Z, Merrifield RB, Schlenstadt G, Zimmermann R (1989) Chemical synthesis and enzymic processing of precursor forms of cecropins A and B. J Biol Chem 264:5852–5860
Chevillard M, Deleage G, Couble P (1986) Amino acid sequence and putative conformational characteristics of the 25 kD silk protein of Bombyx mori. Sericologia 26:435–449
Common IFB (1990) Moths of Australia. Brill, Leiden
Couble P, Moine A, Garel A, Prudhomme JC (1983) Developmental variations of a nonfibroin mRNA of Bombyx mori silkgland, encoding for a low-molecular-weight silk protein. Dev Biol 97:398–407
Gorg A, Postel W, Weser J (1985) Horizontal SDS electrophoresis in ultrathin foregradient gels for the analysis of urinary proteins. Sci Tools (LKB) 32:5–9
Grzelak K, Couble P, Garel A, Kludkiewicz B, Alrouz H (1988) Low molecular weight silk proteins in Galleria mellonella. Insect Biochem 18:223–228
Johnson DA, Gautsch JW, Sportsman JR, Elder JH (1984) Improved technique utilizing nonfat dry milk for analysis of proteins and nucleic acids transferred to nitrocellulose. Gene Anal Tech 1:3–8
Kaufman RJ, Sharp PA (1983) Growth-dependent expression of dihydrofolate reductase mRNA from modular cDNA genes. Mol Cell Biol 3:1598–1608
Kikuchi Y, Mori K, Suzuki S, Yamaguchi K, Mizuno S (1992) Structure of the Bombyx mori fibroin light-chain-encoding gene: upstream sequence elements common to the light and heavy chain. Gene 110:151–158
Kimura K, Oyama F, Ueda H, Mizuno S, Shimura K (1985) Molecular cloning of the fibroin light chain complementary DNA and its use in the study of the expression of the light chain gene in the posterior silk gland of Bombyx mori. Experientia 41:1167–1171
Kodak D (1992) Small protein components of the cocoons in Galleria mellonella (Lepidoptera, Pyralidae) and Bombyx mori (Lepidoptera, Bombycidae). Acta Ent Bohemoslov 89:269–273
Kopáček P, Tesařová Z, Minková Z (1988) Analyses of urine proteins by SDS-polyacrylamide electrophoresis and immunoblotting (in Czech). Biochem Clin Bohemoslov 17:45–54
Kozak M (1989) The scanning model for translation: an update. Mol Cell Biol 9:5134–5142
Kreil G, Haiml L, Suchanek G (1980) Stepwise cleavage of the Pro part of romelittin by dipeptidase IV. Eur J Biochem 111:49–58
Matsudaira P (1987) Sequence from picomole quantities of roteins electroblotted onto polyvinylidine difluoride membranes. J Biol Chem 262:10035–10038
Sasaki T, Noda H (1973) Studies on silk fibroin of Bombyx mori directly extracted from the silk gland. I. Molecular weight determination in guanidine hydrochloride or urea solutions. Biochim Biophys Acta 310:91–103
Sehnal, F. (1966) Kritisches Studium der Bionomie und Biometrik der in verschiedenen Lebensbedingungen geztichteten Wachsmotte, Galleria mellonella L. (Lepidoptera). Z Wiss Zool 174:53–82
Takei F, Kikuchi Y, Kikuchi A, Mizuno S, Shimura K (1987) Further evidence for importance of the subunit combination of silk fibroin in its efficient secretion from the posterior silk gland cells. J Cell Biol 105:175–180
Tanaka K, Mori K, Mizuno S (1993) Immunological identification of the major disulfide-linked light component of silk fibroin. J Biochem. 114:1–4
Von Heijne G (1986) A new method for predicting signal sequence cleavage sites. Nucleic Acids Res 14:4683–4690
Yaffe D, Nudel U, Mayer Y, Neuman S (1985) Highly conserved sequences in the 3′untranslated region of mRNAs coding for homologous proteins in distantly related species. Nucleic Acids Res 13:3723–3737
Yamaguchi K, Kikuchi Y, Takagi T, Kikuchi A, Oyama F, Shimura K., Mizuno S (1989) Primary structure of the silk fibroin light chain determined by cDNA sequencing and peptide analysis. J Mol Biol 210:127–139
Žurovec M, Sehnal F, Scheller K, Kumaran AK (1992) Silk gland specific cDNAs from Galleria mellonella L. Insect Biochem Mol Biol 22:55–67
Author information
Authors and Affiliations
Additional information
Communicated by K. Illmensee
Rights and permissions
About this article
Cite this article
Žurovec, M., Vašková, M., Kodrík, D. et al. Light-chain fibroin of Galleria mellonella L.. Molec. Gen. Genet. 247, 1–6 (1995). https://doi.org/10.1007/BF00425815
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF00425815