RESUMO
In contrast to common meiotic gene conversion, mitotic gene conversion, because it is so rare, is often ignored as a process influencing allelic diversity. We show that if there is a large enough number of premeiotic cell divisions, as seen in many organisms without early germline sequestration, such as plants, this is an unsafe position. From examination of 1.1 million rice plants, we determined that the rate of mitotic gene conversion events, per mitosis, is 2 orders of magnitude lower than the meiotic rate. However, owing to the large number of mitoses between zygote and gamete and because of long mitotic tract lengths, meiotic and mitotic gene conversion can be of approximately equivalent importance in terms of numbers of markers converted from zygote to gamete. This holds even if we assume a low number of premeiotic cell divisions (approximately 40) as witnessed in Arabidopsis. A low mitotic rate associated with long tracts is also seen in yeast, suggesting generality of results. For species with many mitoses between each meiotic event, mitotic gene conversion should not be overlooked.
Assuntos
Conversão Gênica/genética , Variação Genética/genética , Oryza/genética , Plantas/genética , Alelos , Conversão Gênica/fisiologia , Genótipo , Células Germinativas/metabolismo , Meiose/genética , Mitose/genética , Recombinação Genética/genéticaRESUMO
Nature has found many ways to utilize transposable elements (TEs) throughout evolution. Many molecular and cellular processes depend on DNA-binding proteins recognizing hundreds or thousands of similar DNA motifs dispersed throughout the genome that are often provided by TEs. It has been suggested that TEs play an important role in the evolution of such systems, in particular, the rewiring of gene regulatory networks. One mechanism that can further enhance the rewiring of regulatory networks is nonallelic gene conversion between copies of TEs. Here, we will first review evidence for nonallelic gene conversion in TEs. Then, we will illustrate the benefits nonallelic gene conversion provides in rewiring regulatory networks. For instance, nonallelic gene conversion between TE copies offers an alternative mechanism to spread beneficial mutations that improve the network, it allows multiple mutations to be combined and transferred together, and it allows natural selection to work efficiently in spreading beneficial mutations and removing disadvantageous mutations. Future studies examining the role of nonallelic gene conversion in the evolution of TEs should help us to better understand how TEs have contributed to evolution.
Assuntos
Elementos de DNA Transponíveis/genética , Conversão Gênica/genética , Conversão Gênica/fisiologia , Redes Reguladoras de Genes/genética , Redes Reguladoras de Genes/fisiologia , Humanos , Mutação/genéticaRESUMO
Although repair of double-strand breaks (DSBs) by gene conversion is the most accurate way to repair such lesions, in budding yeast there is a 1,000-fold increase in accompanying mutations, including interchromosomal template switches (ICTS) involving highly mismatched (homeologous) ectopic sequences. Although such events are rare and appear at a rate of 2 × 10(-7) when template jumps occur between 71% identical sequences, they are surprisingly frequent (0.3% of all repair events) when the second template is identical to the first, revealing the remarkable instability of repair DNA synthesis. With homeologous donors, ICTS uses microhomologies as small as 2 bp. Cells lacking mismatch repair proteins Msh6 and Mlh1 form chimeric recombinants with two distinct patches of microhomology, implying that these proteins are crucial for strand discrimination of heteroduplex DNA formed during ICTS. We identify the chromatin remodeler Rdh54 as the first protein required for template switching that does not affect simple gene conversion.
Assuntos
Montagem e Desmontagem da Cromatina/genética , Cromossomos Fúngicos , DNA Fúngico/genética , Conversão Gênica/fisiologia , Proteínas de Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/genética , Proteínas Adaptadoras de Transdução de Sinal/genética , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Sequência de Bases , Cromatina/genética , Cromatina/metabolismo , Montagem e Desmontagem da Cromatina/fisiologia , DNA Helicases/genética , DNA Helicases/metabolismo , Reparo de Erro de Pareamento de DNA/genética , Reparo de Erro de Pareamento de DNA/fisiologia , DNA Topoisomerases/genética , DNA Topoisomerases/metabolismo , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Conversão Gênica/genética , Genes Fúngicos/genética , Genes Fúngicos/fisiologia , Genoma Fúngico , Dados de Sequência Molecular , Proteína 1 Homóloga a MutL , Proteínas de Saccharomyces cerevisiae/metabolismo , Homologia de Sequência do Ácido NucleicoRESUMO
La embriogénesis somática es importante como sistema modelo para estudiar el desarrollo de eventos fisiológicos, citológicos y moleculares que sustentan la embriogénesis en plantas, por ser un sistema adecuado para la propagación masiva de especies vegetales y servir de herramienta para el mejoramiento genético, la conservación de germoplasma y la validación de nuevos productos biológicos, y facilitar la producción a gran escala a través del cultivo en medio líquido y su aplicación en biorreactores, proporcionando alta frecuencia de multiplicación, rápido crecimiento del embrión, facilidad de absorción de nutrientes y reducción de la labor de subcultivo. En este trabajo se empleó la embriogénesis somática como vía de multiplicación para evaluar el efecto de metabolitos bacterianos en la inducción de suspensiones celulares y embriones somáticos en tres genotipos de cafeto pertenecientes a Coffea canephora P. variedad Robusta. Para ello se estudiaron densidades de inóculo entre 0,2, 0,5, 1,0 y 3,0 gMF/L-1, y se evaluó el efecto de diferentes medios de cultivo en el desarrollo del proceso. Los resultados mostraron un comportamiento diferenciado en el genotipo M-28, en medios de cultivo suplementados con reguladores de crecimiento convencionales y en los alternativos. Se evidenció una fuerte relación entre la viabilidad celular y el número de células, ante las diferentes condiciones de cultivo y según la densidad de inóculo, se observó un amplio rango de tamaño y forma en las poblaciones de embriones somáticos. Los porcentajes de conversión de ES con el medio MDE-2 evidenciaron mejoras de este indicador para el cultivo del cafeto.
The somatic embryogenesis is important as model system to study the development of physiologic and molecular events that sustain the embryogenesis in plants, is an appropriate system for the massive propagation of vegetable species and as tool for the genetic improvement, the germplasm conservation and the validation of new biological products and to facilitate the multiplication to great scale through the culture in liquid medium, as well as application in bioreactores, providing high multiplication frequency, quick growth of the embryo, easiness of absorption of nutritious and reduction of the subculturing. In this paper the somatic embryogenesis was used to evaluate the effect of bacterial compounds in the induction of cellular suspensions and somatic embryos in three coffee genotypes of Coffea canephora P. var. Robusta. Were studied inoculo densities among 0.2, 0.5, 1.0 and 3.0 gMF/L-1 and the effect of different culture medium in the development of the process. The results showed a behavior differed in the genotype M-28, in medium culture with conventional regulators of growth and the alternatives. Strong relationship was evidenced between the cellular viability and the number of cells, in the different cultivation conditions and according to the inoculo density, a wide range of size and forms as observed in the populations of somatic embryos. The conversion percentages with the medium MDE-2, evidenced improvements of this indicator for the coffee.
Assuntos
Germinação/fisiologia , Germinação/genética , Germinação/imunologia , Conversão Gênica/fisiologia , Conversão Gênica/genética , Conversão Gênica/imunologiaRESUMO
Heterosis is a widespread phenomenon corresponding to the increase in fitness following crosses between individuals from different populations or lines relative to their parents. Its genetic basis has been a topic of controversy since the early 20th century. The masking of recessive deleterious mutations in hybrids likely explains a substantial part of heterosis. The dynamics and consequences of these mutations have thus been studied in depth. Recently, it was suggested that GC-biased gene conversion (gBGC) might strongly affect the fate of deleterious mutations and may have significant fitness consequences. gBGC is a recombination-associated process mimicking selection in favor of G and C alleles, which can interfere with selection, for instance by increasing the frequency of GC deleterious mutations. I investigated how gBGC could affect the amount and genetic structure of heterosis through an analysis of the interaction between gBGC and selection in subdivided populations. To do so, I analyzed the infinite island model both by numerical computations and by analytical approximations. I showed that gBGC might have little impact on the total amount of heterosis but could greatly affect its genetic basis.
Assuntos
Composição de Bases/genética , Composição de Bases/fisiologia , Conversão Gênica/genética , Conversão Gênica/fisiologia , Vigor Híbrido/genética , Vigor Híbrido/fisiologia , Modelos Genéticos , Loci Gênicos/genética , Humanos , Mutação , Seleção GenéticaRESUMO
Exo1 is a member of the Rad2 protein family and possesses both 5'-3' exonuclease and 5' flap endonuclease activities. In addition to performing a variety of functions during mitotic growth, Exo1 is also important for the production of crossovers during meiosis. However, its precise molecular role has remained ambiguous and several models have been proposed to account for the crossover deficit observed in its absence. Here, we present physical evidence that the nuclease activity of Exo1 is essential for normal 5'-3' resection at the Spo11-dependent HIS4 hotspot in otherwise wild-type cells. This same activity was also required for normal levels of gene conversion at the locus. However, gene conversions were frequently observed at a distance beyond that at which resection was readily detectable arguing that it is not the extent of the initial DNA end resection that limits heteroduplex formation. In addition to these nuclease-dependent functions, we found that an exo1-D173A mutant defective in nuclease activity is able to maintain crossing-over at wild-type levels in a number of genetic intervals, suggesting that Exo1 also plays a nuclease-independent role in crossover promotion.
Assuntos
Quebras de DNA de Cadeia Dupla , Reparo do DNA , Exodesoxirribonucleases/fisiologia , Saccharomyces cerevisiae/genética , Oxirredutases do Álcool/genética , Aminoidrolases/genética , Troca Genética , Endodesoxirribonucleases/genética , Exodesoxirribonucleases/genética , Conversão Gênica/genética , Conversão Gênica/fisiologia , Meiose/genética , Meiose/fisiologia , Mutação Puntual , Pirofosfatases/genética , Saccharomyces cerevisiae/enzimologia , Proteínas de Saccharomyces cerevisiae/genéticaRESUMO
BACKGROUND: Horizontal gene transfer (HGT) is relatively common in plant mitochondrial genomes but the mechanisms, extent and consequences of transfer remain largely unknown. Previous results indicate that parasitic plants are often involved as either transfer donors or recipients, suggesting that direct contact between parasite and host facilitates genetic transfer among plants. RESULTS: In order to uncover the mechanistic details of plant-to-plant HGT, the extent and evolutionary fate of transfer was investigated between two groups: the parasitic genus Cuscuta and a small clade of Plantago species. A broad polymerase chain reaction (PCR) survey of mitochondrial genes revealed that at least three genes (atp1, atp6 and matR) were recently transferred from Cuscuta to Plantago. Quantitative PCR assays show that these three genes have a mitochondrial location in the one species line of Plantago examined. Patterns of sequence evolution suggest that these foreign genes degraded into pseudogenes shortly after transfer and reverse transcription (RT)-PCR analyses demonstrate that none are detectably transcribed. Three cases of gene conversion were detected between native and foreign copies of the atp1 gene. The identical phylogenetic distribution of the three foreign genes within Plantago and the retention of cytidines at ancestral positions of RNA editing indicate that these genes were probably acquired via a single, DNA-mediated transfer event. However, samplings of multiple individuals from two of the three species in the recipient Plantago clade revealed complex and perplexing phylogenetic discrepancies and patterns of sequence divergence for all three of the foreign genes. CONCLUSIONS: This study reports the best evidence to date that multiple mitochondrial genes can be transferred via a single HGT event and that transfer occurred via a strictly DNA-level intermediate. The discovery of gene conversion between co-resident foreign and native mitochondrial copies suggests that transferred genes may be evolutionarily important in generating mitochondrial genetic diversity. Finally, the complex relationships within each lineage of transferred genes imply a surprisingly complicated history of these genes in Plantago subsequent to their acquisition via HGT and this history probably involves some combination of additional transfers (including intracellular transfer), gene duplication, differential loss and mutation-rate variation. Unravelling this history will probably require sequencing multiple mitochondrial and nuclear genomes from Plantago. See Commentary: http://www.biomedcentral.com/1741-7007/8/147.
Assuntos
Conversão Gênica/fisiologia , Transferência Genética Horizontal/fisiologia , Genes Mitocondriais/genética , Interações Hospedeiro-Parasita/genética , Plantas/genética , Sequência de Bases , Cuscuta/genética , Cuscuta/fisiologia , Genes de Plantas/fisiologia , Variação Genética/fisiologia , Dados de Sequência Molecular , Filogenia , Desenvolvimento Vegetal , Plantago/genética , Plantago/parasitologia , Pseudogenes , Homologia de Sequência do Ácido NucleicoRESUMO
Distribution of temperature-dependent sex determination (TSD) and genotypic sex determination (GSD) across the phylogeny of dragon lizards implies multiple independent origins of at least one, and probably both, modes of sex determination. Female Pogona vitticeps are the heterogametic sex, but ZZ individuals reverse to a female phenotype at high incubation temperatures. We used reiterated genome walking to extend Z and W chromosome-linked amplified fragment length polymorphism (AFLP) markers, and fluorescence in situ hybridization for physical mapping. One extended fragment hybridized to both W and Z microchromosomes, identifying the Z microchromosome for the first time, and a second hybridized to the centromere of all microchromosomes. W-linked sequences were converted to a single-locus PCR sexing assay. P. vitticeps sex chromosome sequences also shared homology with several other Australian dragons. Further physical mapping and isolation of sex-specific bacterial artificial chromosome clones will provide insight into the evolution of sex determination and sex chromosomes in GSD and TSD dragon lizards.
Assuntos
Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Lagartos/genética , Mapeamento Físico do Cromossomo , Cromossomos Sexuais/genética , Animais , Sequência de Bases , Estruturas Cromossômicas/genética , Cruzamentos Genéticos , Feminino , Conversão Gênica/genética , Conversão Gênica/fisiologia , Cariotipagem , Masculino , Dados de Sequência Molecular , Pseudogenes/genética , Análise para Determinação do SexoRESUMO
Measuring meiotic gene conversion is important both because of its role in the fundamental mechanisms of meiotic recombination and because of its influence on linkage relationships and allelic diversity in the genome. Historically, gene conversion has been most thoroughly examined in fungal organisms through the use of tetrad analysis. Here we describe a method for using tetrad analysis in the model plant Arabidopsis thaliana to detect and quantify gene conversion events - a resource unavailable in most other higher eukaryotic model systems.
Assuntos
Arabidopsis/genética , Biomarcadores/análise , Conversão Gênica/fisiologia , Células Germinativas/metabolismo , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/metabolismo , Biomarcadores/metabolismo , Cromossomos de Plantas , Cruzamentos Genéticos , Células Germinativas/química , Modelos Biológicos , Fenótipo , Plantas Geneticamente ModificadasRESUMO
This study provides the first investigation of the diversity, structure, and molecular evolution of MHII beta genes in a non-model percid species -- the Eurasian perch (Perca fluviatilis L.). PCR primers developed here were highly specific, and documented a high diversity of the MHII beta1 domain in perch. Our results suggest a minimum of eight MHII beta loci in this species -- a finding congruent with several studies suggesting that many Euteleostei posses multiple MHII beta loci. As for other vertebrates, both positive selection and gene-conversion contribute to the reported high allelic diversity. Similarly, the MHII beta1 domain in perch exhibits a characteristic MHC fold known from other vertebrates. In addition, our results suggest some teleost specific differences of the MHII beta1 domain, including: differences in chemical properties of specific amino acids in the beta1 domain, the absence of the tetrapod specific glycolisation signal, and differences in the positions of some of the positively selected codons in the MHII beta1 domain, which are presumably involved in antigen binding. Future studies should investigate the teleost MHII beta genes in more details in order to confirm the suggested differences, and to determine the extent to which these differences prevail in different teleost lineages.
Assuntos
Evolução Molecular , Proteínas de Peixes/genética , Conversão Gênica/fisiologia , Antígenos de Histocompatibilidade Classe II/genética , Percas/genética , Locos de Características Quantitativas/genética , Animais , Sítios de Ligação/fisiologia , Proteínas de Peixes/imunologia , Glicosilação , Antígenos de Histocompatibilidade Classe II/imunologia , Percas/imunologia , Estrutura Terciária de Proteína/fisiologia , Locos de Características Quantitativas/imunologiaRESUMO
Gene conversion, defined as the nonreciprocal transfer of DNA, is one result of homologous recombination. Three steps in recombination could give rise to gene conversion: (i) DNA synthesis for repair of the degraded segment, (ii) Holliday junction migration, leading to heteroduplex formation, and (iii) repair of mismatches in the heteroduplex. There are at least three proteins (RuvAB, RecG, and RadA) that participate in the second step. Their roles have been studied for homologous recombination, but evidence of their relative role in gene conversion is lacking. In this work, we showed the effect on gene conversion of mutations in ruvB, recG, and radA in Rhizobium etli, either alone or in combination, using a cointegration strategy previously developed in our laboratory. The results indicate that the RuvAB system is highly efficient for gene conversion, since its absence provokes smaller gene conversion segments than those in the wild type as well as a shift in the preferred position of conversion tracts. The RecG system possesses a dual role for gene conversion. Inactivation of recG leads to longer gene conversion tracts than those in the wild type, indicating that its activity may hinder heteroduplex extension. However, under circumstances where it is the only migration activity present (as in the ruvB radA double mutant), conversion segments can still be seen, indicating that RecG can also promote gene conversion. RadA is the least efficient system in R. etli but is still needed for the production of detectable gene conversion tracts.
Assuntos
DNA Cruciforme/genética , DNA Cruciforme/fisiologia , Conversão Gênica/genética , Rhizobium etli/genética , Proteínas de Bactérias/genética , Proteínas de Bactérias/fisiologia , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/fisiologia , Conversão Gênica/fisiologia , Modelos GenéticosRESUMO
To maintain genome stability, cells respond to DNA damage by activating signalling pathways that govern cell-cycle checkpoints and initiate DNA repair. Cell-cycle checkpoint controls should connect with DNA repair processes, however, exactly how such coordination occurs in vivo is largely unknown. Here we describe a new role for the E3 ligase RAD18 as the integral component in translating the damage response signal to orchestrate homologous recombination repair (HRR). We show that RAD18 promotes homologous recombination in a manner strictly dependent on its ability to be recruited to sites of DNA breaks and that this recruitment relies on a well-defined DNA damage signalling pathway mediated by another E3 ligase, RNF8. We further demonstrate that RAD18 functions as an adaptor to facilitate homologous recombination through direct interaction with the recombinase RAD51C. Together, our data uncovers RAD18 as a key factor that orchestrates HRR through surveillance of the DNA damage signal.
Assuntos
Quebras de DNA de Cadeia Dupla , Dano ao DNA/fisiologia , Reparo do DNA/fisiologia , Proteínas de Ligação a DNA/fisiologia , Recombinação Genética/fisiologia , Transdução de Sinais/fisiologia , Camptotecina/farmacologia , Linhagem Celular , Cromatina/metabolismo , DNA/efeitos dos fármacos , DNA/metabolismo , DNA/efeitos da radiação , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Conversão Gênica/fisiologia , Células HeLa , Histonas/genética , Histonas/metabolismo , Humanos , Modelos Biológicos , Ligação Proteica/fisiologia , Domínios e Motivos de Interação entre Proteínas/fisiologia , Ubiquitina/metabolismo , Enzimas de Conjugação de Ubiquitina/genética , Ubiquitina-Proteína Ligases , Dedos de Zinco/fisiologiaRESUMO
To cope with an unpredictable variety of potential pathogenic insults, the immune system must generate an enormous diversity of recognition structures, and it does so by making stepwise modifications at key genetic loci in each lymphoid cell. These modifications proceed through the action of lymphoid-specific proteins acting together with the general DNA-repair machinery of the cell. Strikingly, these general mechanisms are usually diverted from their normal functions, being used in rather atypical ways in order to privilege diversity over accuracy. In this Review, we focus on the contribution of a set of DNA polymerases discovered in the past decade to these unique DNA transactions.
Assuntos
Linfócitos B/enzimologia , DNA Polimerase Dirigida por DNA/metabolismo , Imunidade Ativa/fisiologia , Animais , Conversão Gênica/fisiologia , Humanos , Switching de Imunoglobulina/fisiologia , Hipermutação Somática de Imunoglobulina/fisiologiaRESUMO
Gene conversion, one of the two mechanisms of homologous recombination, involves the unidirectional transfer of genetic material from a 'donor' sequence to a highly homologous 'acceptor'. Considerable progress has been made in understanding the molecular mechanisms that underlie gene conversion, its formative role in human genome evolution and its implications for human inherited disease. Here we assess current thinking about how gene conversion occurs, explore the key part it has played in fashioning extant human genes, and carry out a meta-analysis of gene-conversion events that are known to have caused human genetic disease.
Assuntos
Evolução Molecular , Conversão Gênica/fisiologia , Predisposição Genética para Doença , Alelos , Variação Genética , Humanos , Modelos Genéticos , MutaçãoRESUMO
The semen coagulum proteins have undergone substantial structural changes during evolution. In primates, these seminal vesicle-secreted proteins are known as semenogelin I (SEMG1) and semenogelin II (SEMG2). Previous studies on the common marmoset (Callithrix jacchus) showed that ejaculated semen from this New World monkey contains semenogelin, but it remained unclear whether it carries both genes or only SEMG1 and no SEMG2, like the closely related cotton-top tamarin (Saguinus oedipus). In this study we show that there are two genes, both expressed in the seminal vesicles. Surprisingly, the genes show an almost perfect sequence identity in a region of 1.25 kb, encompassing nearly half of the genes and containing exon 1, intron 1, and the first 0.9 kb of exon 2. The underlying molecular mechanism is most likely gene conversion, and a phylogenetic analysis suggests that SEMG1 is the most probable donor gene. The marmoset SEMG1 in this report differs from a previously reported cDNA by a lack of nucleotides encoding one repeat of 60 amino acids, suggesting that marmoset SEMG1 displays allelic size variation. This is similar to what was recently demonstrated in humans, but in marmosets the polymorphism was generated by a repeat duplication, whereas in humans it was a deletion. Together, these studies shed new light on the evolution of semenogelins and the mechanisms that have generated the structural diversity of semen coagulum proteins.
Assuntos
Callithrix/genética , Conversão Gênica/fisiologia , Proteínas Secretadas pela Vesícula Seminal/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Humanos , Masculino , Dados de Sequência Molecular , Filogenia , Homologia de SequênciaRESUMO
Homozygosity or compound heterozygosity for the c.833T>C transition (p.I278 T) in the cystathionine beta-synthase (CBS) gene represents the most common cause of pyridoxine-responsive homocystinuria in Western Eurasians. However, the frequency of the pathogenic c.833C allele, as observed in healthy newborns from several European countries (q(c.833C) approximately equals 3.3 x 10(-3)), is approximately 20-fold higher than expected on the basis of the observed number of symptomatic homocystinuria patients carrying this mutation (q(c.833C) approximately equals 0.18 x 10(-3)), implying clinical underascertainment. Intriguingly, the c.833C mutation is also present in combination with a 68-bp insertion, c.[833C; 844_845ins68], in a substantial proportion of chromosomes from nonhomocystinuric individuals worldwide. We have sought to study the relationship between the pathogenic and nonpathogenic c.833C-bearing chromosomes and to determine whether the pathogenic c.[833C; -] chromosomes are identical-by-descent or instead arose by recurrent mutation. Initial haplotype analysis of 780 randomly selected Czech and sub-Saharan African wild-type chromosomes, employing 12 intragenic markers, revealed 29 distinct CBS haplotypes, of which 10 carried the c.[833C; 844_845ins68] combination; none carried an isolated c.833C or c.844_845ins68 mutation. Subsequent examination of 69 pathogenic c.[833C; -] chromosomes, derived from homocystinuria patients of predominantly European origin, disclosed three unrelated haplotypes that differed from their wild-type counterparts by virtue of the presence of c.833C, thereby indicating that c.833T>C transition has occurred repeatedly and independently in the past. Since c.833T does not reside within an obvious mutational hotspot, we surmise that the three pathogenic and comparatively prevalent c.[833C; -] chromosomes may have originated by recurrent gene conversion employing the common nonpathogenic c.[833C; 844_845ins68] chromosomes as templates.
Assuntos
Cistationina beta-Sintase/genética , Conversão Gênica/fisiologia , Variação Genética , Haplótipos , Homocistinúria/genética , África , Sequência de Bases , Europa (Continente) , Frequência do Gene , Testes Genéticos , Humanos , Dados de Sequência MolecularRESUMO
While evolution of coding sequences has been intensively studied, diversification of noncoding regulatory regions remains poorly understood. In this study, we investigated the molecular evolution of an enhancer region located 5 kb upstream of the transcription start site of the maize pericarp color1 (p1) gene. The p1 gene encodes an R2R3 Myb-like transcription factor that regulates the flavonoid biosynthetic pathway in maize floral organs. Distinct p1 alleles exhibit organ-specific expression patterns on kernel pericarp and cob glumes. A cob glume-specific regulatory region has been identified in the distal enhancer. Further characterization of 6 single-copy p1 alleles, including P1-rr (red pericarp/red cob) and P1-rw (red pericarp and white cob), reveals 3 distinct enhancer types. Sequence variations in the enhancer are correlated with the p1 gene expression patterns in cob glume. Structural comparisons and phylogenetic analyses suggest that evolution of the enhancer region is likely driven by gene conversion between long direct noncoding repeats (approximately 6 kb in length). Given that tandem and segmental duplications are common in both animal and plant genomes, our studies suggest that recombination between noncoding duplicated sequences could play an important role in creating genetic and phenotypic variations.
Assuntos
Conversão Gênica/fisiologia , Genes Reguladores , Variação Genética , Fenótipo , Sequências Repetitivas de Ácido Nucleico , Zea mays/genética , Alelos , Marcadores Genéticos/genética , Dados de Sequência MolecularRESUMO
Mitotic disjunction of the repetitive ribosomal DNA (rDNA) involves specialized segregation mechanisms dependent on the conserved phosphatase Cdc14. The reason behind this requirement is unknown. We show that rDNA segregation requires Cdc14 partly because of its physical length but most importantly because a fraction of ribosomal RNA (rRNA) genes are transcribed at very high rates. We show that cells cannot segregate rDNA without Cdc14 unless they undergo genetic rearrangements that reduce rDNA copy number. We then demonstrate that cells with normal length rDNA arrays can segregate rDNA in the absence of Cdc14 as long as rRNA genes are not transcribed. In addition, our study uncovers an unexpected role for the replication barrier protein Fob1 in rDNA segregation that is independent of Cdc14. These findings demonstrate that highly transcribed loci can cause chromosome nondisjunction.
Assuntos
DNA Ribossômico/genética , Genes de RNAr , Não Disjunção Genética , RNA Ribossômico/biossíntese , Transcrição Gênica/fisiologia , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/fisiologia , Segregação de Cromossomos , Proteínas Fúngicas/genética , Proteínas Fúngicas/fisiologia , Conversão Gênica/fisiologia , Deleção de Genes , Dosagem de Genes , Genes cdc , Modelos Genéticos , Mutação , RNA Polimerase II/metabolismo , Leveduras/citologiaRESUMO
Pseudogenes are remnants of gene duplication (nonprocessed pseudogenes) and retrotransposition (processed pseudogenes) events. This study describes methods for identifying gene conversion candidates from predicted pseudogenes. Pseudogenes may accumulate and harbor sequence variations over time that become disease-causing mutations when transferred to genes by gene conversion. A total of 14,476 pseudogenes were identified, including 3,426 nonprocessed pseudogenes. In addition, 1,945 nonprocessed pseudogenes that are localized near their progenitor gene were evaluated for their possible role in gene conversion and disease. All 11 known, human cases of gene conversion (with deleterious effects) involving pseudogenes were successfully identified by these methods. Among the pseudogenes identified is a retinitis pigmentosa 9 (RP9) pseudogene that carries a c.509A>G mutation which produces a p.Asp170Gly substitution that is associated with the RP9 form of autosomal dominant retinitis pigmentosa (adRP). The c.509A>G mutation in RP9 is a previously unrecognized example of gene conversion between the progenitor gene and its pseudogene. Notably, two processed pseudogenes also contain mutations associated with diseases. An inosine monophosphate dehydrogenase 1 (IMPDH1) pseudogene carries a c.676G>A mutation that produces a p.Asp226Asn substitution that causes the retinitis pigmentosa 10 (RP10) form of adRP; and a phosphoglycerate kinase 1 (PGK1) pseudogene (PGK1P1) carries a c.837T>C mutation that produces a p.Ile252Thr substitution that is associated with a phosphoglycerate kinase deficiency. Ranking of nonprocessed pseudogenes as candidates for gene conversion was also performed based on the sequence characteristics of published cases of pseudogene-mediated gene conversion. All results and tools produced by this study are available for download at: http://genome.uiowa.edu/pseudogenes.
Assuntos
Análise Mutacional de DNA/métodos , Conversão Gênica/fisiologia , Genômica/métodos , Pseudogenes , Biologia Computacional/métodos , Humanos , IMP Desidrogenase/genética , Fosfoglicerato Quinase/deficiência , Proteínas/genética , Fatores de Processamento de RNA , Retinose Pigmentar/genéticaRESUMO
Asexual reproduction is believed to be detrimental, mainly because of the accumulation of deleterious mutations over time, a hypothesis known as Muller's ratchet. In seed plants, most asexually reproducing genetic systems are polyploid, with apomictic species (plants forming seeds without fertilization) as well as plastids and mitochondria providing prominent examples. Whether or not polyploidy helps asexual genetic systems to escape Muller's ratchet is unknown. Gene conversion, particularly when slightly biased, represents a potential mechanism that could allow asexual genetic systems to reduce their mutation load in a genome copy number-dependent manner. However, direct experimental evidence for the operation of gene conversion between genome molecules to correct mutations is largely lacking. Here we describe an experimental system based on transgenic tobacco chloroplasts that allows us to analyze gene conversion events in higher plant plastid genomes. We provide evidence for gene conversion acting as a highly efficient mechanism by which the polyploid plastid genetic system can correct deleterious mutations and make one good genome out of two bad ones. Our finding that gene conversion can be biased may provide a molecular link between asexual reproduction, high genome copy numbers and low mutation rates.