Design of novel peptide inhibitors against the conserved bacterial transcription terminator, Rho
| Autor: | Pankaj Sharma, Ranjan Sen, Amit Kumar, Gairika Ghosh, Sriyans Jain |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Sequence Homology Peptide medicine.disease_cause Biochemistry bacteriophage Bacterial transcription Gene expression ATc anhydrous tetracycline Terminator Regions Genetic EC elongation complex chemistry.chemical_classification Chemistry Escherichia coli Proteins Psu CTD C-terminal domain Terminator (genetics) RB roadblock SBS secondary RNA-binding site Plasmids Protein Binding Research Article PBS primary RNA-binding site Xanthomonas Antimicrobial peptides T-buffer transcription buffer ITC isothermal calorimetry 03 medical and health sciences Peptide Library Rho Escherichia coli medicine Amino Acid Sequence NTD N-terminal domain Molecular Biology transcription termination 030102 biochemistry & molecular biology C-terminus RNA AMP antimicrobial peptides Mycobacterium tuberculosis Cell Biology KFC Knowledge-based FADE and Contacts Peptide Fragments Ni-NTA nickel nitrilotriacetic acid 030104 developmental biology Drug Design peptides Capsid Proteins BSA bovine serum albumin qRT-PCR quantitative reverse transcription PCR |
| Zdroj: | The Journal of Biological Chemistry |
| ISSN: | 0021-9258 |
| DOI: | 10.1016/j.jbc.2021.100653 |
| Popis: | The transcription terminator Rho regulates many physiological processes in bacteria, such as antibiotic sensitivity, DNA repair, RNA remodeling, and so forth, and hence, is a potential antimicrobial target, which is unexplored. The bacteriophage P4 capsid protein, Psu, moonlights as a natural Rho antagonist. Here, we report the design of novel peptides based on the C-terminal region of Psu using phenotypic screening methods. The resultant 38-mer peptides, in addition to containing mutagenized Psu sequences, also contained plasmid sequences, fused to their C termini. Expression of these peptides inhibited the growth of Escherichia coli and specifically inhibited Rho-dependent termination in vivo. Peptides 16 and 33 exhibited the best Rho-inhibitory properties in vivo. Direct high-affinity binding of these two peptides to Rho also inhibited the latter's RNA-dependent ATPase and transcription termination functions in vitro. These two peptides remained functional even if eight to ten amino acids were deleted from their C termini. In silico modeling and genetic and biochemical evidence revealed that these two peptides bind to the primary RNA-binding site of the Rho hexamer near its subunit interfaces. In addition, the gene expression profiles of these peptides and Psu overlapped significantly. These peptides also inhibited the growth of Mycobacteria and inhibited the activities of Rho proteins from Mycobacterium tuberculosis, Xanthomonas, Vibrio cholerae, and Salmonella enterica. Our results showed that these novel anti-Rho peptides mimic the Rho-inhibition function of the ∼42-kDa dimeric bacteriophage P4 capsid protein, Psu. We conclude that these peptides and their C-terminal deletion derivatives could provide a basis on which to design novel antimicrobial peptides. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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