Abstract: The yet uncharacterized membrane protein SA2056 belongs to the ubiquitous RND (Resistance-Nodulation-cell Division) family of transmembrane efflux transporters. The sa2056 gene is located downstream of femX, the gene encoding the essential, non-ribosomal peptidyl-transferase adding the first glycine in the staphylococcal cell wall pentaglycine interpeptide. Due to its proximity to and weak co-transcription with femX, we assumed that sa2056 may somehow be involved in peptidoglycan synthesis. Specific antibodies against SA2056 showed that this protein is expressed during growth and present in the membrane fraction of cell preparations. Using a bacterial two hybrid system, SA2056 was shown to interact (i) with itself, (ii) with FemB, which adds glycines 4 and 5 to the peptidoglycan interpeptide and (iii) with the essential penicillin binding proteins, PBP1 and PBP2, required for cell division and incorporation of the peptidoglycan into the cell wall. Unexpectedly, deletion of sa2056 led to no phenotype regarding growth, antibiotic resistances or cell morphology; nor did sa2056 deletion in combination with femB inactivation alter b-lactam and lysostaphin sensitivity and resistance, respectively, pointing to possible redundancy in the cell wall synthesis pathway. These results suggest an accessory role of SA2056 in S. aureus peptidoglycan synthesis, broadening the range of biological functions of RND proteins.
Keywords: Staphylococcus aureus; RND protein; FemABX; PBP; peptidoglycan; bacterial two-hybrid system
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Quiblier, C.; Luczak-Kadlubowska, A.; Holdener, E.; Alborn, D.; Schneider, T.; Wiedemann, I.; Pinho, M.G.; Sahl, H.-G.; Rohrer, S.; Berger-Bächi, B.; Senn, M.M. The Staphylococcus aureus Membrane Protein SA2056 Interacts with Peptidoglycan Synthesis Enzymes. Antibiotics 2013, 2, 11-27.
Quiblier C, Luczak-Kadlubowska A, Holdener E, Alborn D, Schneider T, Wiedemann I, Pinho MG, Sahl H-G, Rohrer S, Berger-Bächi B, Senn MM. The Staphylococcus aureus Membrane Protein SA2056 Interacts with Peptidoglycan Synthesis Enzymes. Antibiotics. 2013; 2(1):11-27.
Quiblier, Chantal; Luczak-Kadlubowska, Agnieszka; Holdener, Esther; Alborn, Daniela; Schneider, Tanja; Wiedemann, Imke; Pinho, Mariana G.; Sahl, Hans-Georg; Rohrer, Susanne; Berger-Bächi, Brigitte; Senn, Maria M. 2013. " The Staphylococcus aureus Membrane Protein SA2056 Interacts with Peptidoglycan Synthesis Enzymes." Antibiotics 2, no. 1: 11-27.