Article

Molecular cloning and characterization of a cDNA encoding a larval cuticle protein of Bombyx mori.

Department of Biology, Tokyo Metropolitan University, Minamiosawa, Japan.
Comparative Biochemistry and Physiology Part B Biochemistry and Molecular Biology (Impact Factor: 1.9). 02/1999; 122(1):105-9. DOI: 10.1016/S0305-0491(98)10151-7
Source: PubMed

ABSTRACT Cuticle proteins termed LCPs are the major protein components of the larval integument of the silkworm, Bombyx mori. We purified an 18 kDa LCP (LCP18) from the guanidine hydrochloride extract of the larval cuticle and identified an LCP18 cDNA clone. The deduced primary structure and mRNA expression pattern of LCP18 are similar to those of other Bombyx LCPs and to several cuticle proteins identified in other insect species. RNA blot analysis demonstrated that the biosynthesis of LCP18 is regulated in a stage-dependent manner at the level of mRNA in epidermal cells. An in vivo study using a juvenile hormone analogue suggested that juvenile hormone positively regulates expression of LCP18 mRNA during larval intermolt stages.

0 Followers
 · 
81 Views
  • [Show abstract] [Hide abstract]
    ABSTRACT: BMCP18(2) is one of the major cuticle proteins identified in the larval cuticle of the silkworm, Bombyx mori. A genomic clone coding for BMCP18 was isolated from a B. mori genomic library, and its structure was analyzed. The BMCP18 gene consists of three exons interspersed by two introns. Bm1 element-like sequences were identified around this gene, suggesting possible involvement of this retroposon in the duplication of B. mori cuticle protein genes during evolution. A structural comparison of the BMCP18 gene and related cuticle protein genes of other lepidopteran species (MSCP14.6 and HCCP12) showed that the 5' upstream region of the BMCP18, MSCP14.6, and HCCP12 genes has a 12-bp identical sequence matching the recognition sequence for transcription factors COUP-TF and HNF-4. This implies that molecular mechanisms regulating expression of these cuticle protein genes are also conserved. mRNAs coding for Bmsvp, the B. mori homolog of Drosophila Seven-up, which is known as a homolog of vertebrate COUP-TF, and BmHNF-4, a homolog of vertebrate HNF-4, were detected in the larval epidermis. Bmsvp bound to the 12-bp sequence in vitro, suggesting that Bmsvp regulates the BMCP18 gene expression.
    Insect Biochemistry and Molecular Biology 05/2001; 31(6-7):611-20. DOI:10.1016/S0965-1748(00)00166-1 · 3.42 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: The wing-deficient mutant, fl├╝gellos (fl), of the silkworm lacks four wings in the pupa and the adult, due to aberrant wing morphogenesis during metamorphosis. To elucidate the mechanisms of wing-specific deficiencies in the fl mutant, we used mRNA differential display and identified five genes abnormally expressed in the fl wing discs. Northern blot and RT-PCR analyses revealed that four genes were overexpressed, but the fifth one was not transcribed in the fl wing discs. The expression level of ribosome-associated protein p40 in the fl wing discs was elevated approximately 10 times compared to the wild-type (WT) discs. Another overexpressed gene CB10 encodes a novel wing-specific protein with a putative zinc-finger motif. Overexpression of two components of extracellular matrix, cuticle protein 18 (BMCP18) and a fibrillin-like protein AD10, may result in the abnormal wing morphogenesis in the fl mutant. In contrast, a novel member of multifunctional Ca2+-binding protein annexins, designated as annexin b13 (Anx b13), was expressed dominantly in the wing discs of WT but completely repressed in the fl tissues. Strong expression of Anx b13 in wing discs during the fourth and fifth instar indicates that ANX B13 plays an important role in wing morphogenesis.
    Insect Biochemistry and Molecular Biology 07/2002; 32(6):691-9. DOI:10.1016/S0965-1748(01)00148-5 · 3.42 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: When insects molt, the exoskeleton is renewed under the controls of insect hormones via the biosynthesis and degradation of cuticle proteins. To understand the hormonal control of cuticle formation, we used the differential display method to look for stage-specific cuticle genes, and identified a novel cDNA named Bombyx mori Cuticle Protein GlyGlyTyr-repeat 1 (BMCPG1). Expression of BMCPG1 mRNA peaked sharply immediately after a pulse of ecdysteroid during the fourth molt and pre-pupal stages, concurrent with the expression of genes for FTZF1 and dopa decarboxylase. BMCPG1 was expressed only in the epidermis, but not in any other tissue. We cultured the larval epidermis and found that BMCPG1 expression is not induced by the continuous presence of ecdysteroid. Removal of ecdysteroid from the medium, which constitutes a pulse treatment, is required for the induction of BMCPG1 transcription. These results explain well the stage-specific expression of BMCPG1 by ecdysteroid in vivo. Based on its expression patterns and unique structure, we propose that BMCPG1 may be a novel component of epicuticle of B. mori, and is probably involved in cross-linking of proteins via its GGY repeats.
    Insect Biochemistry and Molecular Biology 07/2002; 32(6):599-607. DOI:10.1016/S0965-1748(01)00136-9 · 3.42 Impact Factor