RESUMO
The form of RNA processing known as SL trans-splicing involves the transfer of a short conserved sequence, the spliced leader (SL), from a noncoding SL RNA to the 5' ends of mRNA molecules. SL trans-splicing occurs in several animal taxa, including bdelloid rotifers (Rotifera, Bdelloidea). One striking feature of these aquatic microinvertebrates is the large proportion of foreign genes, i.e. those acquired by horizontal gene transfer from other organisms, in their genomes. However, whether such foreign genes behave similarly to native genes has not been tested in bdelloids or any other animal. We therefore used a combination of experimental and computational methods to examine whether transcripts of foreign genes in bdelloids were SL trans-spliced, like their native counterparts. We found that many foreign transcripts contain SLs, use similar splice acceptor sequences to native genes, and are able to undergo alternative trans-splicing. However, a significantly lower proportion of foreign mRNAs contains SL sequences than native transcripts. This demonstrates a novel functional difference between foreign and native genes in bdelloids and suggests that SL trans-splicing is not essential for the expression of foreign genes, but is acquired during their domestication.
Assuntos
Transferência Genética Horizontal , Genoma Helmíntico , RNA de Helmintos/genética , RNA Mensageiro/genética , RNA Líder para Processamento/genética , Rotíferos/genética , Trans-Splicing , Processamento Alternativo , Sequência de Aminoácidos , Animais , Evolução Biológica , Perfilação da Expressão Gênica , Ontologia Genética , Anotação de Sequência Molecular , RNA de Helmintos/metabolismo , RNA Mensageiro/metabolismo , RNA Líder para Processamento/metabolismo , Alinhamento de Sequência , Transcriptoma , TransgenesRESUMO
The immunodominant lipopolysaccharide is a key antigenic factor for Gram-negative pathogens such as salmonellae where it plays key roles in host adaptation, virulence, immune evasion, and persistence. Variation in the lipopolysaccharide is also the major differentiating factor that is used to classify Salmonella into over 2600 serovars as part of the Kaufmann-White scheme. While lipopolysaccharide diversity is generally associated with sequence variation in the lipopolysaccharide biosynthesis operon, extraneous genetic factors such as those encoded by the glucosyltransferase (gtr) operons provide further structural heterogeneity by adding additional sugars onto the O-antigen component of the lipopolysaccharide. Here we identify and examine the O-antigen modifying glucosyltransferase genes from the genomes of Salmonella enterica and Salmonella bongori serovars. We show that Salmonella generally carries between 1 and 4 gtr operons that we have classified into 10 families on the basis of gtrC sequence with apparent O-antigen modification detected for five of these families. The gtr operons localize to bacteriophage-associated genomic regions and exhibit a dynamic evolutionary history driven by recombination and gene shuffling events leading to new gene combinations. Furthermore, evidence of Dam- and OxyR-dependent phase variation of gtr gene expression was identified within eight gtr families. Thus, as O-antigen modification generates significant intra- and inter-strain phenotypic diversity, gtr-mediated modification is fundamental in assessing Salmonella strain variability. This will inform appropriate vaccine and diagnostic approaches, in addition to contributing to our understanding of host-pathogen interactions.
Assuntos
Antígenos de Bactérias/genética , Transferência Genética Horizontal/genética , Glicosiltransferases/genética , Interações Hospedeiro-Patógeno/imunologia , Salmonella enterica/patogenicidade , Antígenos de Bactérias/metabolismo , Variação Genética , Genoma Bacteriano , Bactérias Gram-Negativas/genética , Interações Hospedeiro-Patógeno/genética , Humanos , Lipopolissacarídeos/genética , Salmonella enterica/genéticaRESUMO
An improved method of non-radioactive identification of transcription start sites is presented in which the use of 7-deaza dGTP in the primer extension reaction allows the product to be directly aligned to cycle sequencing traces on an automated sequencer. This removes the documented need to apply corrections for mobility differences.
Assuntos
Biologia Molecular/métodos , Sítio de Iniciação de Transcrição , Automação/métodos , Pareamento de Bases , Desoxiguanosina/análogos & derivados , Desoxiguanosina/metabolismo , Análise de Sequência de DNA/métodosRESUMO
The Caulobacter crescentus DNA adenine methyltransferase CcrM and its homologs in the alpha-Proteobacteria are essential for viability. CcrM is 34% identical to the yhdJ gene products of Escherichia coli and Salmonella enterica. This study provides evidence that the E. coli yhdJ gene encodes a DNA adenine methyltransferase. In contrast to an earlier report, however, we show that yhdJ is not an essential gene in either E. coli or S. enterica.
