Journal: Nucleic Acids Research

Article Title: PhoP-regulated VirK acts as an accessory factor to maintain virulence in polymyxin-resistant Klebsiella pneumoniae

doi: 10.1093/nar/gkag290

Figure Lengend Snippet: Transcriptomic analysis of Mut-S in comparison to Kpn2146 strains. ( A ) Volcano map displaying the upregulated (red dots) and downregulated (yellow dots) genes between the Mut-S group and the control group. ( B ) Changes in the relative expression of the mgrB, phoP , and virK genes in the Mut-S strain compared with those in the WT strain. ( C )Changes in the virK gene expression in the Mut-S strain after supplementation with the mgrB gene. ( D ) Changes in phoP and virK gene expressions after phoP gene knockdown in the Mut-S strain. ( E ) Changes in the mgrB and virK gene expressions after the mgrB gene was complemented in the phoP knockout strain in Mut-S. ( F ) Changes in the phoP and virK gene expressions after the phoP gene was supplemented in the phoP knockout strains in Mut-S. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: The effects of virK on bacterial pathogenicity were further evaluated in a mouse systemic infection model using virK mutants and complemented strains under K. pneumoniae ATCC BAA2146 (wild type) or Mut-S ( mgrB truncated strain) backgrounds.

Techniques: Comparison, Control, Expressing, Gene Expression, Knockdown, Knock-Out

Journal: iScience

Article Title: A mechanism of target mRNA selection and activity regulation in meiosis-related RBM46-MEIOC-YTHDC2 complex

doi: 10.1016/j.isci.2026.116234

Figure Lengend Snippet: Determination of RBM46-MEIOC-YTHDC2-targeting motif in the Rad21 and Meioc 3′UTR (A) Diagram of firefly luciferase reporter containing the 3′UTR of interest. This backbone vector contains poly(A) signals downstream of firefly luciferase reporter gene. (B) Schematic illustration of the workflow of the luciferase experiment. (C) Full-length and truncated Rad21 3′UTR constructs (ΔF1 to ΔF7). (D) Dual luciferase activities of the indicated constructs (ΔF1 to ΔF4) were measured 48 h after transfection, N = 3. (E) Relative luciferase activities of the indicated constructs (ΔF5 to ΔF7), N = 3. (F) Full-length and truncated Meioc 3′UTR constructs (ΔD1 to ΔD7). (G) Relative luciferase activities from the indicated constructs (ΔD1 to ΔD4), N = 3. (H) Relative luciferase activities of the indicated constructs (ΔD5 to ΔD7), N = 3. (I) Consensus sequence of ΔF5, ΔF7, and ΔD5 fragments was analyzed using the STREME algorithm. This sequence contains RBM46 binding motif AAUCAU identified by eCLIP using mouse testes and is similar to the consensus sequence identified by an in vitro RBM46 binding assay. (J) Mut-ΔF5 construct were generated by deletion of consensus sequence in ΔF5. Compared to ΔF5, the luciferase activities were rescued in cells transfected with Mut-ΔF5, N = 3. (K) Relative luciferase activities from Mut-ΔD5 construct were derepressed, N = 3. (L) Relative luciferase activities from Mut- Actb were repressed by RMY complex. N = 3. Data are presented as mean ± SEM. p values were determined by one-way ANOVA with Benjamini-Hochberg correction.

Article Snippet: The Mut-ΔF5 and Mut-ΔD5 sequences were synthesized from Sangon Biotech and then inserted into pmirGLO vector.

Techniques: Luciferase, Plasmid Preparation, Construct, Transfection, Sequencing, Binding Assay, In Vitro, Generated

Journal: Nucleic Acids Research

Article Title: PhoP-regulated VirK acts as an accessory factor to maintain virulence in polymyxin-resistant Klebsiella pneumoniae

doi: 10.1093/nar/gkag290

Figure Lengend Snippet: Transcriptomic analysis of Mut-S in comparison to Kpn2146 strains. ( A ) Volcano map displaying the upregulated (red dots) and downregulated (yellow dots) genes between the Mut-S group and the control group. ( B ) Changes in the relative expression of the mgrB, phoP , and virK genes in the Mut-S strain compared with those in the WT strain. ( C )Changes in the virK gene expression in the Mut-S strain after supplementation with the mgrB gene. ( D ) Changes in phoP and virK gene expressions after phoP gene knockdown in the Mut-S strain. ( E ) Changes in the mgrB and virK gene expressions after the mgrB gene was complemented in the phoP knockout strain in Mut-S. ( F ) Changes in the phoP and virK gene expressions after the phoP gene was supplemented in the phoP knockout strains in Mut-S. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: During preliminary investigations of a polymyxin-resistant mutant (Mut-S) of K. pneumoniae ATCC BAA-2146 (Kpn2146) [ ], we detected highly upregulated expression of the Kpn2146:RS17285 gene.

Techniques: Comparison, Control, Expressing, Gene Expression, Knockdown, Knock-Out