RESUMO
Defensins are small cationic peptides involved in innate immunity and are components of the first line of defence against invading pathogens. beta-defensins are a subgroup of the defensin family that display a particular cysteine spacing and pattern of intramolecular bonding. These molecules are produced mostly by epithelia lining exposed surfaces and appear to have both antimicrobial and cell signalling functions. The unusually high degree of sequence variation in the mature peptide produced by the paralogous and in some cases orthologous genes implies extensive specialisation and species specific adaptation. Here we review recent functional data that are an important addition to our knowledge of the innate immune response and novel antibiotic design. We also consider the organisation and evolution of the genomic loci harbouring these genes where radical and rapid changes in beta-defensin sequences have been shown to result from the interplay of both positive and negative selection. Consequently these genes provide some unusually clear glimpses of the processes of duplication and specialisation that have shaped the mammalian genome.
Assuntos
Evolução Molecular , Imunidade Inata/genética , Seleção Genética , beta-Defensinas/genética , Sequência de Aminoácidos , Animais , Sítios de Ligação , Cromossomos de Mamíferos , Genoma , Humanos , Camundongos , Dados de Sequência Molecular , Filogenia , Homologia de Sequência de Aminoácidos , Relação Estrutura-Atividade , alfa-Defensinas/genética , beta-Defensinas/fisiologiaRESUMO
Chromosome deletions in the mouse have proven invaluable in the dissection of gene function. The brown deletion complex comprises >28 independent genome rearrangements, which have been used to identify several functional loci on chromosome 4 required for normal embryonic and postnatal development. We have constructed a 172-bacterial artificial chromosome contig that spans this 22-megabase (Mb) interval and have produced a contiguous, finished, and manually annotated sequence from these clones. The deletion complex is strikingly gene-poor, containing only 52 protein-coding genes (of which only 39 are supported by human homologues) and has several further notable genomic features, including several segments of >1 Mb, apparently devoid of a coding sequence. We have used sequence polymorphisms to finely map the deletion breakpoints and identify strong candidate genes for the known phenotypes that map to this region, including three lethal loci (l4Rn1, l4Rn2, and l4Rn3) and the fitness mutant brown-associated fitness (baf). We have also characterized misexpression of the basonuclin homologue, Bnc2, associated with the inversion-mediated coat color mutant white-based brown (B(w)). This study provides a molecular insight into the basis of several characterized mouse mutants, which will allow further dissection of this region by targeted or chemical mutagenesis.
Assuntos
Deleção Cromossômica , Glicoproteínas de Membrana/genética , Oxirredutases/genética , Animais , Sequência de Bases , Evolução Biológica , Mapeamento Cromossômico , Cromossomos Artificiais Bacterianos/genética , Feminino , Morte Fetal/genética , Genes Letais , Cor de Cabelo/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Mutantes , Fenótipo , Polimorfismo de Nucleotídeo Único , GravidezRESUMO
PURPOSE: The present study examined the developmental and tissue expression of the retinitis pigmentosa GTPase regulator (RPGR) gene in Xenopus laevis. METHODS: The cDNA for X. laevis RPGR (XRPGR) was isolated from adult eye mRNA by reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends. The deduced peptide sequence was aligned with RPGR orthologues. Gene expression was examined by whole-mount in situ hybridization and RT-PCR. The localization of XRPGR in X. laevis photoreceptor cells and XTC-2 cells was determined by immunostaining. RESULTS: The XRPGR(ex1-19) isoform encodes a protein of 727 amino acids containing an RCC1 domain and a C-terminal isoprenylation anchorage motif. It shares 33% to 41% amino acid identity with human, mouse, and dog RPGR. The C-terminal exon of the alternatively spliced RPGR(ORF15) isoform is also conserved across species. XRPGR is expressed at the earliest stages of X. laevis development and persists into adulthood, where expression is highest in the eye. XRPGR is expressed in presumptive eye fields (stages 18 to 22), becoming largely restricted to the central retina (stages 28 to 40). XRPGR protein colocalizes with beta-tubulin at the X. laevis ciliary axoneme and with gamma-tubulin at centrosomes in XTC-2 cells. CONCLUSIONS: XRPGR is widely expressed throughout development but shows highest expression after the appearance of the eye primordium and persists in the eye into adulthood. The data are consistent with XRPGR expression in a single microtubular organelle-the centriole or basal body and associated ciliary transitional zone found in modified sensory cilia of photoreceptors and motile cilia.
Assuntos
Embrião não Mamífero/metabolismo , Proteínas do Olho/genética , Regulação da Expressão Gênica no Desenvolvimento , Fatores de Troca do Nucleotídeo Guanina/genética , Células Fotorreceptoras de Vertebrados/metabolismo , Proteínas de Xenopus/genética , Sequência de Aminoácidos , Animais , Linhagem Celular , Centrossomo/metabolismo , Clonagem Molecular , DNA Complementar/análise , Proteínas do Olho/metabolismo , Técnica Indireta de Fluorescência para Anticorpo , Fatores de Troca do Nucleotídeo Guanina/metabolismo , Hibridização In Situ , Dados de Sequência Molecular , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNA , Tubulina (Proteína)/metabolismo , Proteínas de Xenopus/metabolismo , Xenopus laevis/embriologiaRESUMO
Slow-twitch muscle fibers of the zebrafish myotome undergo a unique set of morphogenetic cell movements. During embryogenesis, slow-twitch muscle derives from the adaxial cells, a layer of paraxial mesoderm that differentiates medially within the myotome, immediately adjacent to the notochord. Subsequently, slow-twitch muscle cells migrate through the entire myotome, coming to lie at its most lateral surface. Here we examine the cellular and molecular basis for slow-twitch muscle cell migration. We show that slow-twitch muscle cell morphogenesis is marked by behaviors typical of cells influenced by differential cell adhesion. Dynamic and reciprocal waves of N-cadherin and M-cadherin expression within the myotome, which correlate precisely with cell migration, generate differential adhesive environments that drive slow-twitch muscle cell migration through the myotome. Removing or altering the expression of either protein within the myotome perturbs migration. These results provide a definitive example of homophilic cell adhesion shaping cellular behavior during vertebrate development.