Molecular characterization of a novel patched-related protein in Apis mellifera and Drosophila melanogaster.
The molecular identification and characterization of the patched-related (ptr) gene and protein in Apis mellifera and Drosophila melanogaster are reported. Ptr proteins are closely related in predicted topology and domain organization to the protein encoded by the Drosophila segment polarity gene patched. Ptrs have 12 potential transmembrane domains arranged in two sets of 1+5 membrane-spanning segments containing a conserved sterol-sensing domain (SSD) and functional GxxxD and PPXY motifs. Phylogenetic analysis showed that Ptrs belong to a previously uncharacterized class of insect proteins that share a high level of sequence identity. Analysis using quantitative real-time polymerase chain reaction (qPCR) indicates that ptr gene is preferentially expressed during embryo stages of A. mellifera development; interestingly, this pattern of temporal expression was also observed for the D. melanogaster homologue, suggesting that these proteins might be involved in embryo morphogenesis. To understand Ptr function at the molecular level, we investigated the subcellular distribution of DmPtr. We have shown by biochemical analysis that DmPtr protein is tightly associated with membranes. Consistently, Ptr immunoreactivity appears to be localized at the sites of membrane furrow formation during cellularization of D. melanogaster embryos. These studies indicated that Ptrs belong to a previously uncharacterized class of insect transmembrane proteins that share a high level of sequence identity. Our analysis of ptr gene expression and protein localization suggest that Ptr might fulfil a developmental role by participating in processes that require growth and stabilization of plasma membrane.
Laboratorio de Bioinformática y Expresión Génica, INTA, Universidad de Chile, Macul 5540, CP 138-11, Santiago, Chile.
Luis
L
Pastenes
Freddy
F
Ibáñez
Carmen
C
Bolatto
Leonardo
L
Pavéz
Verónica
V
Cambiazo
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001516:Bees / Q000235:genetics* / Q000378:metabolism; D015152:Blotting, Northern; D003001:Cloning, Molecular; D029721:Drosophila Proteins / Q000235:genetics* / Q000378:metabolism; D004331:Drosophila melanogaster / Q000235:genetics* / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D007150:Immunohistochemistry; D019476:Insect Proteins / Q000235:genetics* / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D010802:Phylogeny; D012313:RNA / Q000737:chemistry / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D013347:Subcellular Fractions / Q000378:metabolism; D014162:Transfection
D029721:Drosophila Proteins; D019476:Insect Proteins; D008565:Membrane Proteins; D012313:RNA