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 KHdomain protein (KhpA). DkhpA suppresses deletions in most, but not all (mltG), genes involved in peripheral PG synthesis and in the gpsB regulatory gene. DkhpA mutations reduce growth rate, decrease cell size, minimally affect shape and induce expression of the WalRK cell-wall stress regulon. Reciprocal coimmunoprecipitations show that KhpA forms a complex in cells with another KH-domain protein (KhpB/ JAG/EloR). DkhpA and DkhpB 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 Dpbp2b 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 Dpbp2b. However, increased FtsA only partially accounts for the phenotypes of DkhpA mutants. Together, these results suggest that multimeric KhpA/B may function as a pleiotropic RNA chaperone controlling pneumococcal cell division.

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. This article is protected by copyright. All rights reserved.

Absence of the KhpA and KhpB (JAG/EloR) RNA-Binding Proteins Suppresses the Requirement for PBP2b by Overproduction of FtsA in Streptococcus pneumoniae D39 / Zheng Jiaqi, J; Perez Amilcar, J; Tsui Ho ching, Tiffany; Massidda, Orietta; Winkler Malcolm, E.. - In: MOLECULAR MICROBIOLOGY. - ISSN 0950-382X. - 106:5(2017), pp. 793-814. [10.1111/mmi.13847]

Absence of the KhpA and KhpB (JAG/EloR) RNA-Binding Proteins Suppresses the Requirement for PBP2b by Overproduction of FtsA in Streptococcus pneumoniae D39

Massidda, Orietta;
2017-01-01

Abstract

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. This article is protected by copyright. All rights reserved.
2017
5
Zheng Jiaqi, J; Perez Amilcar, J; Tsui Ho ching, Tiffany; Massidda, Orietta; Winkler Malcolm, E.
Absence of the KhpA and KhpB (JAG/EloR) RNA-Binding Proteins Suppresses the Requirement for PBP2b by Overproduction of FtsA in Streptococcus pneumoniae D39 / Zheng Jiaqi, J; Perez Amilcar, J; Tsui Ho ching, Tiffany; Massidda, Orietta; Winkler Malcolm, E.. - In: MOLECULAR MICROBIOLOGY. - ISSN 0950-382X. - 106:5(2017), pp. 793-814. [10.1111/mmi.13847]
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