RESUMO
The transposons of Drosophila melanogaster are regulated by small RNAs that interact with the Piwi family of proteins. These piRNAs are generated from transposons inserted in special loci such as the telomere-associated sequences at the left end of the X chromosome. Drosophila's P transposons can also be regulated by a polypeptide encoded by the KP element, a 1.15-kb-long member of the P family. Using piRNA-generating telomeric P elements (TPs) and repressor-producing transgenic KP elements, we demonstrate a functional connection between these two modes of regulation. By themselves, the TPs partially repress gonadal dysgenesis, a trait caused by rampant P-element activity in the germ line. This repression is manifested as a strictly maternal effect arising from the cytoplasmic transmission of P-specific piRNAs from mother to offspring. The repression is enhanced by genetic interactions between the TPs and other, non-telomeric P elements-a phenomenon attributable to ping-pong amplification of maternal piRNAs. KP elements, like other kinds of non-telomeric P elements, enhance regulation anchored in the TPs. However, with some TPs, the enhanced regulation is manifested as a strictly zygotic effect of the KP element. This effect is seen when the TP has few sequences in common with the KP element, a condition not conducive to ping-pong amplification of piRNAs; it can be attributed to the action of the KP repressor polypeptide. Because the effect is seen only when a TP was present in the mother's genotype, maternally generated P-element piRNAs could facilitate regulation by the KP repressor polypeptide.
Assuntos
Elementos de DNA Transponíveis/genética , Drosophila/genética , Disgenesia Gonadal/genética , Hibridização Genética , Peptídeos/metabolismo , RNA Interferente Pequeno/metabolismo , Animais , Animais Geneticamente Modificados , Cruzamentos Genéticos , Feminino , Padrões de Herança/genética , Masculino , Telômero/metabolismo , TransgenesRESUMO
The X-linked telomeric P elements (TPs) TP5 and TP6 regulate the activity of the entire P element family because they are inserted in a major locus for the production of Piwi-interacting RNAs (piRNAs). The potential for this cytotype regulation is significantly strengthened when either TP5 or TP6 is combined with a non-telomeric X-linked or autosomal transgene that contains a P element. By themselves, none of the transgenic P elements have any regulatory ability. Synergism between the telomeric and transgenic P elements is much greater when the TP is derived from a female. Once an enhanced regulatory state is established in a female, it is transmitted to her offspring independently of either the telomeric or transgenic P elements - that is, it works through a strictly maternal effect. Synergistic regulation collapses when either the telomeric or the transgenic P element is removed from the maternal genotype, and it is significantly impaired when the TPs come from stocks heterozygous for mutations in the genes aubergine, piwi or Su(var)205. The synergism between telomeric and transgenic P elements is consistent with a model in which P piRNAs are amplified by alternating, or ping-pong, targeting of primary piRNAs to sense and antisense P transcripts, with the sense transcripts being derived from the transgenic P element and the antisense transcripts being derived from the TP.
Assuntos
Elementos de DNA Transponíveis/genética , Drosophila melanogaster/genética , Genes Ligados ao Cromossomo X , Telômero/genética , Transgenes/genética , Animais , Animais Geneticamente Modificados , Homólogo 5 da Proteína Cromobox , Elementos de DNA Transponíveis/fisiologia , Proteínas de Drosophila , Drosophila melanogaster/citologia , Drosophila melanogaster/metabolismo , Feminino , Regulação da Expressão Gênica , Genes de Insetos , Genes Ligados ao Cromossomo X/genética , RNA Interferente Pequeno/genéticaRESUMO
TP5, a P element inserted in the telomere-associated sequences of the X chromosome, represses the excision of other P elements in the germ line through a combination of maternal and zygotic effects. The maternal component of this repression is impaired by heterozygous mutations in the aubergine and Suppressor of variegation 205 genes; one mutation in the piwi gene also appears to impair repression. In the female germ line, the level of TP5 mRNA is increased by these impairing mutations. The impairing aubergine and piwi mutations also increase the level of germ-line mRNA from CP, a transgene that encodes the P-element transposase; however, the Suppressor of variegation 205 mutation does not. These findings are discussed in terms of a model of P-element regulation that involves post-transcriptional and chromatin re-organizing events mediated by maternally transmitted small RNAs derived from the telomeric P element.
Assuntos
Proteínas Cromossômicas não Histona/genética , Elementos de DNA Transponíveis/genética , Drosophila melanogaster/genética , Genes de Insetos , Animais , Proteínas Argonautas , Homólogo 5 da Proteína Cromobox , Proteínas de Drosophila/genética , Proteínas do Ovo/genética , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Células Germinativas , Masculino , Mutação/genética , Fatores de Iniciação de Peptídeos , Complexo de Inativação Induzido por RNA/genética , Telômero/genética , Transposases/genética , Cromossomo X/genética , Cromossomo X/fisiologiaRESUMO
Strains carrying the X-linked telomeric P elements TP5 or TP6 varied in their ability to repress hybrid dysgenesis. The rank ordering of these strains was consistent across different genetic assays and was not related to the type of telomeric P element (TP5 or TP6) present. Strong repression of dysgenesis was associated with weak expression of mRNA from the telomeric P element and also with a reduced amount of mRNA from a transposase-producing P element contained within a transgene inserted on an autosome. A strictly maternal component of repression, transmitted independently of the telomeric P element, was detected in the daughters but not the sons of females from the strongest repressing strains. However, this effect was seen only when dysgenesis was induced by crossing these females to males from a P strain, not when it was induced by crossing them to males homozygous for a single transposase-producing P element contained within a transgene. These findings are consistent with the hypothesis that the P cytotype, the condition that regulates P elements, involves an RNA interference mechanism mediated by piRNAs produced by telomeric P elements such as TP5 and TP6 and amplified by RNAs produced by other P elements.
Assuntos
Regulação da Expressão Gênica , Genes Ligados ao Cromossomo X , Telômero/genética , Transposases , Animais , Cruzamentos Genéticos , Elementos de DNA Transponíveis/genética , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , Feminino , Genes de Insetos , Disgenesia Gonadal , Masculino , Interferência de RNA , Transgenes , Transposases/genética , Transposases/metabolismoRESUMO
The X-linked telomeric P elements TP5 and TP6 interact synergistically with non-telomeric P elements to repress hybrid dysgenesis. In this repression, the telomeric P elements exert maternal effects, which, however, are not sufficient to establish synergism with the non-telomeric P elements. Once synergism is established, the capacity to repress dysgenesis in the offspring of a cross persists for at least two generations after removing the telomeric P element from the genotype. At the molecular level, synergism between telomeric and non-telomeric P elements is correlated with effective elimination of P-element mRNA in the germ line. Maternally transmitted mutations in the genes aubergine, piwi and Suppressor of variegation 205 [Su(var)205] block the establishment of synergism between telomeric and non-telomeric P elements, and paternally transmitted mutations in piwi and Su(var)205 disrupt synergism that has already been established. These findings are discussed in terms of a model of cytotype regulation of P elements based on Piwi-interacting RNAs (piRNAs) that are amplified by cycling between sense and antisense species.
Assuntos
Elementos de DNA Transponíveis/genética , Drosophila melanogaster/genética , Regulação da Expressão Gênica , Telômero/genética , Animais , Animais Geneticamente Modificados , Homólogo 5 da Proteína Cromobox , Cruzamentos Genéticos , Proteínas de Drosophila , Genes de Insetos , RNA Mensageiro/metabolismoRESUMO
Transposons, especially retrotransposons, are abundant in the genome of Drosophila melanogaster. These mobile elements are regulated by small RNAs that interact with the Piwi family of proteins-the piwi-interacting or piRNAs. The Piwi proteins are encoded by the genes argonaute3 (ago3), aubergine (aub), and piwi. Heterochromatin Protein 1 (HP1), a chromatin-organizing protein encoded by the Suppressor of variegation 205 [Su(var)205] gene, also plays a role in this regulation. To assess the mutational impact of weakening the system for transposon regulation, we measured the frequency of recessive X-linked lethal mutations occurring in the germ lines of males from stocks that were heterozygous for mutant alleles of the ago3, aub, piwi, or Su(var)205 genes. These mutant alleles are expected to deplete the wild-type proteins encoded by these genes by as much as 50%. The mutant alleles of piwi and Su(var)205 significantly increased the X-linked lethal mutation frequency, whereas the mutant alleles of ago3 did not. An increased mutation frequency was also observed in males from one of two mutant aub stocks, but this increase may not have been due to the aub mutant. The increased mutation frequency caused by depleting Piwi or HP1suggests that chromatin-organizing proteins play important roles in minimizing the germ-line mutation rate, possibly by stabilizing the structure of the heterochromatin in which many transposons are situated.
Assuntos
Elementos de DNA Transponíveis , Drosophila melanogaster/genética , Mutação em Linhagem Germinativa , RNA Interferente Pequeno/fisiologia , Animais , Proteínas Argonautas/genética , Homólogo 5 da Proteína Cromobox , Proteínas Cromossômicas não Histona/genética , Proteínas de Drosophila/genética , Masculino , Fatores de Iniciação de Peptídeos/genéticaRESUMO
Previous studies have shown that telomeric P elements inserted at the left end of the X chromosome are anchors of the P cytotype, the maternally inherited state that regulates P-element activity in the germ line of Drosophila melanogaster. This regulation is mediated by small RNAs that associate with the Piwi family of proteins (piRNAs). We extend the analysis of cytotype regulation by studying new combinations of telomeric and nontelomeric P elements (TPs and non-TPs). TPs interact with each other to enhance cytotype regulation. This synergism involves a strictly maternal effect, called presetting, which is apparently mediated by piRNAs transmitted through the egg. Presetting by a maternal TP can elicit regulation by an inactive paternally inherited TP, possibly by stimulating its production of primary piRNAs. When one TP has come from a stock heterozygous for a mutation in the aubergine, piwi, or Suppressor of variegation 205 genes, the synergism between two TPs is impaired. TPs also interact with non-TPs to enhance cytotype regulation, even though the non-TPs lack regulatory ability on their own. Non-TPs are not susceptible to presetting by a TP, nor is a TP susceptible to presetting by a non-TP. The synergism between TPs and non-TPs is stronger when the TP was inherited maternally. This synergism may be due to the accumulation of secondary piRNAs created by ping-pong cycling between primary piRNAs from the TPs and mRNAs from the non-TPs. Maternal transmission of P-element piRNAs plays an important role in the maintenance of strong cytotype regulation over generations.