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The Bacillus subtilis class A penicillin-binding protein 4 (PBP4) requires an accessory protein RpdA

Ruoqi Huang1, Yesha Patel1, John D Helmann1

  • 1Department of Microbiology, Cornell University, Ithaca, NY 14853-8101, USA.

Biorxiv : the Preprint Server for Biology
|January 7, 2026
PubMed

Insights

RpdA is a newly identified regulator of PBP4 in Bacillus subtilis, crucial for cell wall synthesis. Loss of RpdA enhances resistance to cefuroxime by affecting PBP4 localization and activity.

Area of Science:

  • Microbiology
  • Bacterial Cell Wall Synthesis
  • Protein Regulation

Background:

  • Peptidoglycan (PG) synthesis is vital for bacterial cell integrity.
  • Class A and B penicillin-binding proteins (PBPs) orchestrate PG synthesis for growth and division in Bacillus subtilis.
  • PBP4 is a class A PBP involved in PG synthesis.

Purpose of the Study:

  • To identify regulators of PBP4 function in Bacillus subtilis.
  • To elucidate the role of RpdA (formerly YufK) in PBP4 activity and localization.
  • To understand the impact of RpdA on bacterial response to antibiotics.

Main Methods:

  • Genetic analysis, including gene inactivation and epistasis studies.
  • Antibiotic sensitivity assays using cefuroxime (CEF).
  • Cellular localization studies of PBP4.

Main Results:

  • RpdA was identified as a specific regulator of PBP4.
  • Loss of RpdA confers resistance to cefuroxime, similar to PBP4 inactivation.
  • RpdA is essential for the membrane localization and activity of PBP4.
  • RpdA may also play a role in undecaprenyl-phosphate recycling.

Conclusions:

  • RpdA acts as a PBP4 accessory protein, critical for its localization and function.
  • RpdA is involved in bacterial cell wall integrity and antibiotic resistance.
  • The findings reveal a novel regulatory mechanism in bacterial PG synthesis.

Keywords:
PBP4Peptidoglycancefuroximepenicillin-binding proteinsregulator

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