RESUMO
One of the rewards of having a Drosophila melanogaster whole-genome sequence will be the potential to understand the molecular bases for structural features of chromosomes that have been a long-standing puzzle. Analysis of 2.6 megabases of sequence from the tip of the X chromosome of Drosophila identifies 273 genes. Cloned DNAs from the characteristic bulbous structure at the tip of the X chromosome in the region of the broad complex display an unusual pattern of in situ hybridization. Sequence analysis revealed that this region comprises 154 kilobases of DNA flanked by 1.2-kilobases of inverted repeats, each composed of a 350-base pair satellite related element. Thus, some aspects of chromosome structure appear to be revealed directly within the DNA sequence itself.
Assuntos
Drosophila melanogaster/genética , Cromossomo X/genética , Animais , Bandeamento Cromossômico , Biologia Computacional , Cosmídeos , Elementos de DNA Transponíveis , DNA Satélite , Genes de Insetos , Hibridização In Situ , Sequências Repetitivas de Ácido Nucleico , Análise de Sequência de DNA , Cromossomo X/ultraestruturaRESUMO
Leydig insulin-like protein (Ley I-L) is a novel member of the insulin-like hormone superfamily. We report here the isolation and expression of the mouse Ley I-L gene. The gene encodes a polypeptide of 122 amino acids that shows a relatively weak homology (54%) to human and porcine prepro-Ley I-L. However, the predicted B and A chain of the mature mouse Ley I-L exhibit similarities of 73% and 71% with human and porcine Ley I-L, respectively. Alignment of the 5' flanking region of the mouse gene with those of human and porcine did not exhibit any significant sequence homology. However, it contains the conserved sequence of the Ad4 binding site that is present in all promoter regions of steroidogenic P-450 genes and the Müllerian inhibitor substance gene and is recognized by steroidogenic factor 1. The Ley I-L gene is expressed at a high level in the testis and at a much lower level in the ovary. No transcripts could be detected in placenta prepared between days 10 and 19 of pregnancy. Ley I-L transcripts were first detected in fetal testis at 13.5 dpc. After birth, transcript levels remain constant during the following 3 weeks, increasing at the stage in which the first wave of round spermatids undergo spermiogenesis suggesting a functional role of the Ley I-L in early stages of spermatogenesis and germ-cell maturation. In the ovary, the expression of Ley I-L was first detected at day 6 after birth. The pattern of Ley I-L expression at various stages of the estrous cycle and during pregnancy showed a correlation with follicle development.
Assuntos
Regulação da Expressão Gênica no Desenvolvimento , Ovário/metabolismo , Proteínas/genética , Testículo/metabolismo , Sequência de Aminoácidos , Animais , Sequência de Bases , Northern Blotting , Clonagem Molecular , Primers do DNA , Feminino , Insulina , Masculino , Camundongos , Camundongos Endogâmicos , Dados de Sequência Molecular , Ovário/embriologia , Reação em Cadeia da Polimerase , Gravidez , Proteínas/química , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Mapeamento por Restrição , Análise de Sequência , Homologia de Sequência de Aminoácidos , Testículo/embriologiaRESUMO
We present the sequence of a contiguous 2.63 Mb of DNA extending from the tip of the X chromosome of Drosophila melanogaster. Within this sequence, we predict 277 protein coding genes, of which 94 had been sequenced already in the course of studying the biology of their gene products, and examples of 12 different transposable elements. We show that an interval between bands 3A2 and 3C2, believed in the 1970s to show a correlation between the number of bands on the polytene chromosomes and the 20 genes identified by conventional genetics, is predicted to contain 45 genes from its DNA sequence. We have determined the insertion sites of P-elements from 111 mutant lines, about half of which are in a position likely to affect the expression of novel predicted genes, thus representing a resource for subsequent functional genomic analysis. We compare the European Drosophila Genome Project sequence with the corresponding part of the independently assembled and annotated Joint Sequence determined through "shotgun" sequencing. Discounting differences in the distribution of known transposable elements between the strains sequenced in the two projects, we detected three major sequence differences, two of which are probably explained by errors in assembly; the origin of the third major difference is unclear. In addition there are eight sequence gaps within the Joint Sequence. At least six of these eight gaps are likely to be sites of transposable elements; the other two are complex. Of the 275 genes in common to both projects, 60% are identical within 1% of their predicted amino-acid sequence and 31% show minor differences such as in choice of translation initiation or termination codons; the remaining 9% show major differences in interpretation.