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Even small wall-less bacteria have specially developed macro-molecular structures to support their survival. <jats:italic>Mycoplasma pneumoniae<\/jats:italic>, a human pathogen, forms a polar extension called an attachment organelle, which mediates cell division, cytadherence, and cell movement at host cell surface. This characteristic ultrastructure has a constant size of 250\u2013300 nm, but its design principle remains unclear. In this study, we constructed several mutants by genetic manipulation to increase or decrease coiled-coil regions of HMW2, a major component protein of 200 kDa aligned in parallel along the cell axis. HMW2-engineered mutants produced both long and short attachment organelles, which we quantified by transmission electron microscopy and fluorescent microscopy with nano-meter precision. This simple design of HMW2 acting as a molecular ruler for the attachment organelle should provide an insight into bacterial cellular organization and its function for their parasitic lifestyles.<\/jats:p>","DOI":"10.1371\/journal.ppat.1009621","type":"journal-article","created":{"date-parts":[[2021,6,10]],"date-time":"2021-06-10T23:27:40Z","timestamp":1623367660000},"page":"e1009621","update-policy":"https:\/\/doi.org\/10.1371\/journal.ppat.corrections_policy","source":"Crossref","is-referenced-by-count":14,"title":["Molecular ruler of the attachment organelle in Mycoplasma pneumoniae"],"prefix":"10.1371","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8201-2608","authenticated-orcid":true,"given":"Daisuke","family":"Nakane","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kohki","family":"Murata","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3198-4263","authenticated-orcid":true,"given":"Tsuyoshi","family":"Kenri","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1233-6842","authenticated-orcid":true,"given":"Keigo","family":"Shibayama","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takayuki","family":"Nishizaka","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2021,6,10]]},"reference":[{"issue":"6280","key":"ppat.1009621.ref001","doi-asserted-by":"crossref","first-page":"aad6253","DOI":"10.1126\/science.aad6253","article-title":"Design and synthesis of a minimal bacterial genome","volume":"351","author":"CA Hutchison","year":"2016","journal-title":"Science"},{"issue":"21","key":"ppat.1009621.ref002","doi-asserted-by":"crossref","first-page":"3675","DOI":"10.1128\/JB.01865-14","article-title":"Mycoplasma pneumoniae, an underutilized model for bacterial cell biology","volume":"196","author":"MF Balish","year":"2014","journal-title":"J Bacteriol"},{"issue":"2","key":"ppat.1009621.ref003","doi-asserted-by":"crossref","first-page":"579","DOI":"10.1128\/IAI.01036-13","article-title":"In vitro spatial and temporal analysis of Mycoplasma pneumoniae colonization of human airway epithelium","volume":"82","author":"OA Prince","year":"2014","journal-title":"Infect Immun"},{"issue":"1","key":"ppat.1009621.ref004","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1111\/j.1365-2958.2006.05507.x","article-title":"Cytoskeletal protein P41 is required to anchor the terminal organelle of the wall-less prokaryote Mycoplasma pneumoniae","volume":"63","author":"BM Hasselbring","year":"2007","journal-title":"Mol Microbiol"},{"issue":"1","key":"ppat.1009621.ref005","doi-asserted-by":"crossref","first-page":"519","DOI":"10.1146\/annurev.micro.112408.134116","article-title":"Unique centipede mechanism of Mycoplasma gliding","volume":"64","author":"M. 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