Assembly properties of bacterial actin MreB involved in Spiroplasma swimming motility

• Published in: The Journal of biological chemistry Vol. 299; no. 6; p. 104793
• Main Authors: Takahashi, Daichi, Miyata, Makoto, Fujiwara, Ikuko
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.06.2023; American Society for Biochemistry and Molecular Biology
• Subjects: actin; Actins - metabolism; ATPase; bacteria; Bacterial Proteins - metabolism; bundle formation; cell motility; cytoskeleton; Cytoskeleton - metabolism; electron microscopy; Mollicutes; Mycoplasma; polymerization dynamics; Spiroplasma - metabolism; Swimming; TmMreB; bacteria; cytoskeleton; ATPase; EM; Mycoplasma; SpeMreB5; SpeMreB3; CcMreB; cell motility; Mollicutes; actin; SciMreB5; polymerization dynamics; EcMreB; electron microscopy; bundle formation
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/j.jbc.2023.104793

Bacterial actin MreB forms filaments composed of antiparallel double-stranded units. The wall-less helical bacterium Spiroplasma has five MreB homologs (MreB1–5), some of which are involved in an intracellular ribbon for driving the bacterium’s swimming motility. Although the interaction between MreB units is important for understanding Spiroplasma swimming, the interaction modes of each ribbon component are unclear. Here, we examined the assembly properties of Spiroplasma eriocheiris MreB5 (SpeMreB5), one of the ribbon component proteins that forms sheets. Electron microscopy revealed that sheet formation was inhibited under acidic conditions and bundle structures were formed under acidic and neutral conditions with low ionic strength. We also used solution assays and identified four properties of SpeMreB5 bundles as follows: (I) bundle formation followed sheet formation; (II) electrostatic interactions were required for bundle formation; (III) the positively charged and unstructured C-terminal region contributed to promoting lateral interactions for bundle formation; and (IV) bundle formation required Mg2+ at neutral pH but was inhibited by divalent cations under acidic pH conditions. During these studies, we also characterized two aggregation modes of SpeMreB5 with distinct responses to ATP. These properties will shed light on SpeMreB5 assembly dynamics at the molecular level.