Absence of the KhpA and KhpB (JAG/EloR) RNA‐binding proteins suppresses the requirement for PBP2b by overproduction of FtsA in Streptococcus pneumoniae D39

Summary Suppressor mutations were isolated that obviate the requirement for essential PBP2b in peripheral elongation of peptidoglycan from the midcells of dividing Streptococcus pneumoniae D39 background cells. One suppressor was in a gene encoding a single KH‐domain protein (KhpA). ΔkhpA suppresses deletions in most, but not all (mltG), genes involved in peripheral PG synthesis and in the gpsB regulatory gene. ΔkhpA mutations reduce growth rate, decrease cell size, minimally affect shape and induce expression of the WalRK cell‐wall stress regulon. Reciprocal co‐immunoprecipitations show that KhpA forms a complex in cells with another KH‐domain protein (KhpB/JAG/EloR). ΔkhpA and ΔkhpB mutants phenocopy each other exactly, consistent with a direct interaction. RNA‐immunoprecipitation showed that KhpA/KhpB bind an overlapping set of RNAs in cells. Phosphorylation of KhpB reported previously does not affect KhpB function in the D39 progenitor background. A chromosome duplication implicated FtsA overproduction in Δpbp2b suppression. We show that cellular FtsA concentration is negatively regulated by KhpA/B at the post‐transcriptional level and that FtsA overproduction is necessary and sufficient for suppression of Δpbp2b. However, increased FtsA only partially accounts for the phenotypes of ΔkhpA mutants. Together, these results suggest that multimeric KhpA/B may function as a pleiotropic RNA chaperone controlling pneumococcal cell division. RNA binding proteins KhpA and KhpB (JAG; EloR), which are virulence factors, form a complex in cells of the Gram‐positive pathogen, Streptococcus pneumoniae. KhpA/B negatively regulates ftsA expression post‐transcriptionally, and FtsA overproduction in ΔkhpA/B mutants is necessary and sufficient for suppression of some, but not all, mutations in certain essential genes that mediate peptidoglycan synthesis. The combined results of this study suggest that multimeric KhpA/B may act as a pleiotropic RNA chaperone controlling cell division.