Poster abstracts

Poster number 65 submitted by Anne Witzky

Multi-step 5-aminopentanol assembly on Elongation factor P influences antibiotic resistance in Bacillus subtilis

Anne Witzky (Department of Molecular Genetics, Ohio State University, Columbus, Ohio), Rodney Tollerson II (Department of Microbiology, Ohio State University, Columbus, Ohio), Katherine R. Hummels (Department if Biology, Indiana University, Bloomington, Indiana), Andrei Rajkovic (Molecular Cellular Developmental Biology Program, Ohio State University, Columbus Ohio), Daniel B. Kearns (Department if Biology, Indiana University, Bloomington, Indiana), Michael Ibba (Department of Microbiology, Ohio State University, Columbus, Ohio)

Abstract:
Elongation Factor P (EF-P) is a universally conserved translation factor that facilitates translation of polyproline motifs. In order to perform this function, EF-P requires a post-translational modification (PTM) on a conserved residue. Although the position of the modification is highly conserved, the structure can vary widely between organisms. In Bacillus subtilis, EF-P is modified at Lysine 32 with a 5-aminopentanol moiety. However, the modification pathway for this PTM is not known. Here, we use a forward genetic screen to identify genes required for 5-aminopentanolylation. When each gene is deleted, EF-P retains a unique incomplete modification, indicating that 5-aminopentanol is likely directly assembled on EF-P in a multi-step manner. In the presence of each incomplete modification, EF-P activity is impaired to varying degrees. In further phenotypic characterization of these mutants, we found that a ∆efp strain was resistant to sulfonamide drugs and puromycin. The modification mutants had varying degrees of intermediate phenotypes. These findings not only establish a novel EF-P PTM pathway, but they also highlight the importance of EF-P in antibiotic resistance.

References:
Rajkovic A, Hummels KR, Witzky A, Erickson S, Gafken PR, et al. (2016) Translation control
of swarming proficiency in Bacillus subtilis by 5-amino-pentanolylated elongation factor
P. J Biol Chem. 291(21):10976-85.

Keywords: Elongation Factor P, Translation , Post Translational Modification