Bacteriophage T7 terminator T is a class I intrinsic terminator coding for an RNA hairpin structure immediately followed by oligo(U), which has been extensively studied in terms of its transcription termination mechanism, but little is known about its physiological or regulatory functions. In this study, using a T7 mutant phage, where a 31-bp segment of T was deleted from the genome, we discovered that deletion of T from T7 reduces the phage burst size but delays lysis timing, both of which are disadvantageous for the phage. The burst downsizing could directly result from T deletion-caused upregulation of gene 17.5, coding for holin, among other T downstream genes, because infection of gp17.5-overproducing Escherichia coli by wild-type T7 phage showed similar burst downsizing. However, the lysis delay was not associated with cellular levels of holin or lysozyme or with rates of phage adsorption. Instead, when allowed to evolve spontaneously in five independent adaptation experiments, the Tlacking mutant phage, after 27 or 29 passages, recovered both burst size and lysis time reproducibly by deleting early genes 0.5, 0.6, and 0.7 of class I, among other mutations. Deletion of genes 0.5 to 0.7 from the T-lacking mutant phage decreased expression of several T downstream genes to levels similar to that of the wild-type phage. Accordingly, phage T7 lysis timing is associated with cellular levels of T downstream gene products. This suggests the involvement of unknown factor(s) besides the known lysis proteins, lysozyme and holin, and that T plays a role of optimizing burst size and lysis time during T7 infection. IMPORTANCEBacteriophages are bacterium-infecting viruses. After producing numerous progenies inside bacteria, phages lyse bacteria using their lysis protein(s) to get out and start a new infection cycle. Normally, lysis is tightly controlled to ensure phage progenies are maximally produced and released at an optimal time. Here, we have discovered that phage T7, besides employing its known lysis proteins, additionally uses its transcription terminator T to guarantee the optimal lysis of the E. coli host. T, positioned in the middle of the T7 genome, must be inactivated at least partially to allow for transcription-driven translocation of T7 DNA into hosts and expression of T downstream but promoter-lacking genes. What role is played by T before inactivation? Without T, not only was lysis time delayed but also the number of progenies was reduced in this study. Furthermore, T7 can overcome T deletion by further deleting some genes, highlighting that a phage has multiple strategies for optimizing lysis. Bacteriophage T7 is an obligate lytic Escherichia coli phage that has been extensively studied for more than 60 years (1). T7 RNA polymerase, the only RNA polymerase (gp1) in the T7 genome, is one of the best-characterized RNA polymerases, and its transcription mechanisms have been studied in detail (2-6). In the T7 genome, an intrinsic termination signal for T7 RNA polymerase, terminator T, is positioned ...

Single-stranded microRNAs (miRNAs) regulate gene expression by triggering mRNA degradation and/or inhibiting mRNA translation. miRNAs play important roles in various critical cellular processes and are associated with numerous human diseases, including cancer and neurodegenerative diseases. miRNA sequences are embedded in the primary miRNA transcripts (pri-miRNAs) that are initially processed by the Microprocessor complex in the nucleus. Microprocessor can orient itself on pri-miRNAs in two ways: one orientation results in subsequent miRNA production, and the other leads to cleavage of the miRNA sequence. Therefore, orienting Microprocessor on pri-miRNAs is a fundamental mechanism for determining the accuracy and efficiency of pri-miRNA processing and, in turn, miRNA production. Multiple mechanisms controlling Microprocessor orientation on pri-miRNAs, involving both cis-acting RNA elements and trans-acting factors, have recently been revealed. In this review, we discuss these exciting mechanisms and consider potential unknown mechanisms that might regulate Microprocessor orientation on pri-miRNAs.

Background Invasive meningococcal disease (IMD) persists in military units in Vietnam despite the availability of antibiotics and vaccines. A hindrance to reducing the incidence of IMD in Vietnam is a lack of molecular data from isolates of the causative agent, Neisseria meningitidis from this country. Here, we characterized key genetic and epidemiological features of an invasive N. meningitidis isolate from a military unit in Vietnam using whole-genome sequencing. Methods Neisseria meningitidis was isolated from a conscript admitted for meningitis and tested against seven antibiotics. DNA from the isolate was extracted and sequenced using the Illumina HiSeq platform. Denovo assembly and scaffolding were performed to construct a draft genome assembly, from which genes were predicted and functionally annotated. Genome analysis included epidemiological characterization, genomic composition and identification of antibiotic resistance genes. Results Susceptibility testing of the isolate showed high levels of resistance to chloramphenicol and diminished susceptibility to ampicillin and rifampicin. A draft genome of ~ 2.1 Mb was assembled, containing 2451 protein coding sequences, 49 tRNAs and 3 rRNAs. Fifteen coding sequences sharing ≥ 84% identity with known antibiotic resistance genes were identified. Genome analysis revealed abundant repetitive DNAs and two prophages. Epidemiological typing revealed newly described sequence type, antigenic finetype and Bexsero ® Antigen Sequence Typing (BAST). The BAST profile showed no coverage by either Bexsero ® or Trumenba ® . Conclusions Our results present the first genome assembly of an invasive N. meningitidis isolate from a military unit in Vietnam. This study illustrates the usefulness of whole genome sequencing (WGS) analysis for epidemiological and antibiotic resistance studies and surveillance of IMD in Vietnam.

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A large-scale food poisoning outbreak happened at a school canteen in Ninh Binh Province, Vietnam, in 2018, resulting in the hospitalization of 352 students with clinical symptoms indicative of a staphylococcal food poisoning. A subsequent laboratory investigation detected Staphylococcus aureus in two food items—deep-fried shrimp and chicken floss—at up to 103 CFU/mL, and staphylococcal enterotoxins (SEs) in chicken floss at ≥ 0.211 ng SEs/g. S. aureus was also isolated from patients’ vomit and stool samples, and kitchen workers’ stool samples, as well as in frozen chicken meat, but not on the kitchen workers’ hand surfaces, suggesting the cause of this food poisoning outbreak was S. aureus contamination of the chicken meat. Molecular characterization revealed the S. aureus strains isolated from all samples were closely related; all belonged to sequence type (ST) ST6 and spa type t701 and carried both sea and sec genes. This SE-producing strain was resistant to penicillin and tetracycline, while still susceptible to oxacillin, erythromycin, gentamicin, methicillin, and vancomycin. Since S. aureus food poisonings are often underreported, our investigation added to the sparse qualitative and quantitative data of pathogenic S. aureus monitoring and surveillance in Vietnam, providing needed knowledge to guide preventative measures for future outbreaks.