Project description:Genome-wide transcriptional responses of Yersinia entomophaga to shifts in temperature (25 - 37 degrees C) and in vitro and in vivo growth conditions were investigated using RNA-seq.
Project description:Identification the proteins co-secreted with YenTc and compare them to those remaining in yersinia entomophaga cells by bottom-up proteomics.
Project description:Yersinia entomophaga MH96 is a highly virulent entomopathogen that infects pest insects of economic importance. We discovered a new regulator, Yen6, which influences the virulence of MH96 in Galleria mellonella at 37 ˚C. Yen6 is directly upstream of the yen7 gene, which encodes for a RoeA-like PhoB-containing regulator and that is directly upstream of the Yen Toxin component (Yen-Tc) genes. We found that yen6 is the most temperature-responsive gene in MH96, producing both messenger RNA and non-coding RNA in its 3' untranslated region (UTR). Yen6 encodes a unique LytTR-containing transcription factor that targets sugar uptake genes and an RNA-binding protein. Of interest, the non-coding RNA from the yen6 3’ UTR overlaps with yen7 in the opposite orientation. Deleting yen6 did not significantly affect the transcriptional activity of yen7 or downstream Yen-Tc genes during infection at 37 ˚C. However, when we deleted the yen6 locus or induced yen6 plus a 478-bp region of its 3’ UTR at 25 ˚C, we observed a significant increase in the transcription/ translation rates of the Yen-Tc gene chi1 using a chi1:lacZ fusion. Our findings reveal a novel mode of virulence regulation in MH96, where the yen6 gene exerts control over multiple genes involved in G. mellonella infection and exhibits potential for dual functionality at both transcriptional and translational levels.
Project description:Investigation of whole genome gene expression level changes in Yersinia intermedia strain ATCC 29909 in response to oxygen. The experiments and results have not been published yet (manuscript has been submitted to journal office and is under revision)
Project description:Here we report the draft genome of Yersinia entomophaga type strain MH96T. The genome shows 93.8% nucleotide sequence identity to that of Yersinia nurmii type strain APN3a-cT, and comprises a single chromosome of approximately 4,275,531 bp. In silico analysis identified that, in addition to the previously documented Y. entomophaga Yen-TC gene cluster, the genome encodes a diverse array of toxins, including two type III secretion systems, and five rhs-associated gene clusters. As well as these multicomponent systems, several orthologs of known insect toxins, such as VIP2 toxin and the binary toxin PirAB, and distant orthologs of some mammalian toxins, including repeats-in-toxin, a cytolethal distending toxin, hemolysin-like genes and an adenylate cyclase were identified. The genome also contains a large number of hypothetical proteins and orthologs of known effector proteins, such as LopT, as well as genes encoding a wide range of proteolytic determinants, including metalloproteases and pathogen fitness determinants, such as genes involved in iron metabolism. The bioinformatic data derived from the current in silico analysis, along with previous information on the pathobiology of Y. entomophaga against its insect hosts, suggests that a number of these virulence systems are required for survival in the hemocoel and incapacitation of the insect host.
Project description:Purpose: In previous studies, we found that the fyuA gene plays an important role in the virulence and pathogenicity of Yersinia pestis strain 201. In order to globally observe which functions of the fyuA gene also affect Y. pestis, we performed RNA-seq on the Y. pestis wild strain 201-WT and mutant strains △fyuA and △fyuAGCAdel, hoping to find their differences at the transcription level, so that better elucidate the effect of fyuA gene on Y. pestis gene transcription. Methods: Total RNA was extracted using the PureLink™ RNA Mini Kit, and then used for creating a cDNA library and deep sequencing. According to the values of Fragments Per Kilobase of Transcript per Million Mapped Reads (FPKM), the ratio of transcript levels between WT and ΔfyuAGCAdel groups was used as the logarithm to the base 2 (twofold change). The differential values of at least a 2-fold were applied to analyze the differential expression of genes according to the Y. pestis 91001 genome annotation. The transcriptome data were verified by qPCR. Results: We found that compared with 201-WT, genes related to siderophore synthesis such as ybtS, ybtX, ybtQ, irp2, irp1, ybtU, ybtT and ybtE were significantly down-regulated at either 26 ℃ or 37 ℃. At 37 ℃, the genes related to type Ⅲ secretion system were significantly up-regulated.
