Research Projects

Rationale: Carbapenem-resistant hypermucoid hypervirulent Klebsiella pneumoniae is responsible for causing both community-acquired and nosocomial infections, leading to high mortality worldwide. Foundational research into the functional significance of uncharacterized membrane and cellular proteins, including unprecedented signal transduction elements, is imperative, not only to uncover previously overlooked yet crucial roles in metabolism, membrane integrity, virulence, and AMR, but also to identify their pleiotropic potential as validated drug targets. Objective: The objective of this research proposal is to elucidate the Molecular, Biochemical, Biological, Regulatory functions of the Novel Phage-Encoded Eukaryotic-like Serine/Threonine Kinase – Phosphatase Regulatory Module in Hypervirulent Klebsiella pneumoniae. Endogenous substrate identification, cross talk amongst signaling proteins and non-canonical signaling paradigm will be explored for the very first time. Hypothesis: We hypothesize that a novel phage-encoded eukaryotic-like serine/threonine kinase–phosphatase system plays a critical regulatory role in K. pneumoniae by modulating its ability to sense, adapt, and respond to physiological stress and environmental cues. This regulatory mechanism may underlie the pathogen's dual success as both a highly transmissible community-acquired agent and a foremost cause of hospital-acquired infections, characteristics that have placed K. pneumoniae on the WHO's list of highest-priority pathogens. Methodology: Biochemical Characterization of the Novel Phage-Origin Ser/Thr Kinase, Domain Mapping and Motif Analysis to find Conserved and Essential Residues, Characterization of the Hypothetical Serine/Threonine Metallo-Phosphatase 1 in K. pneumoniae. Functional Characterization in Clonally Distinct Isolates through knock-out/complementation studies, Clinical Relevance in Dominantly Circulating Sequence Types, Transcriptomic and Proteomic Approaches to Uncover New Signaling Paradigms. Implications of phosphor Signaling on Effector Proteins and Its Impact on Membrane Integrity, Stress Response, and Klebsiella Biology and Screening for PhagSTKKP antagonist in CR-hmhvKP: Significance and Novelty: For the very first time, a novel phage-encoded eukaryotic-like serine/threonine kinase and its corresponding phosphatase will be characterized in Klebsiella pneumoniae, specifically in a hypervirulent, sequence type (ST)-specific lineage. This kinase uniquely contains a LapB domain at its C-terminal—such a kinase has never been identified or characterized in bacteria before. We are the first team to report and investigate this unprecedented protein. An enigmatic, hierarchical regulatory system governing LPS synthesis, virulence, metabolism, antimicrobial resistance (AMR), and the emergence, transmission, and persistence will be elucidated for the first time in bacteria, using K. pneumoniae as a model organism.