Characterization of holins, the membrane proteins of coliphage ASEC2201: a genomewide in silico approach

作者：Humaira Saeed, Sudhaker Padmesh, Aditi Singh, Sujeet Pratap Singh, Mohammed Haris Siddiqui, Manodeep Sen, Imran Hussain, Mirza Masroor Ali Beg · 发表于：Frontiers in Microbiology · 年份：2025 · DOI：10.3389/fmicb.2025.1550594 · 被引用次数：4 · 研究领域：Bacteriophages and microbial interactions、RNA modifications and cancer、RNA and protein synthesis mechanisms

Drug-resistant Escherichia coli poses a significant healthcare burden, driving the search for novel antimicrobials. We have previously done the isolation and whole-genome sequencing of ASEC2201, a novel coliphage derived from multidrug-resistant clinical E. coli strains. Here, we report the identification and characterization of phage enzyme, holin by in silico approaches. Genome annotation using Prokka identified three putative holin genes (PROKKA_03659, PROKKA_04292, and PROKKA_04422) belonging to the Phage_holin_2_1 superfamily. Upstream promoter prediction revealed active regulatory elements at positions 112, 177, and 186 for these genes, indicating robust transcriptional activity. Transmembrane topology analysis using DeepTMHMM confirmed the presence of two to three α -helical membrane-spanning domains in each holin, essential for pore formation. Homology modeling with SWISS-MODEL yielded high-confidence three-dimensional structures characterized by conserved membrane-anchoring motifs, as supported by QMEAN and GMQE quality scores. In silico identification of cell-penetrating peptide motifs within the holin sequences suggests potential for enhanced intracellular delivery in CPP-fusion therapeutic constructs. Overall, our in-depth analysis elucidates the structural and functional properties of ASEC2201 holins, underscoring their biotechnological significance as scaffolds for developing novel antimicrobial strategies against MDR E. coli . It gives us an understanding on ho...