Cargando…
Cross-kymography analysis to simultaneously quantify the function and morphology of the archaellum
In many microorganisms helical structures are important for motility, e.g., bacterial flagella and kink propagation in Spiroplasma eriocheiris. Motile archaea also form a helical-shaped filament called the ‘archaellum’ that is functionally equivalent to the bacterial flagellum, but structurally rese...
Autores principales: | , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Biophysical Society of Japan (BSJ)
2018
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6018435/ https://www.ncbi.nlm.nih.gov/pubmed/29955563 http://dx.doi.org/10.2142/biophysico.15.0_121 |
_version_ | 1783334949313576960 |
---|---|
author | Kinosita, Yoshiaki Nishizaka, Takayuki |
author_facet | Kinosita, Yoshiaki Nishizaka, Takayuki |
author_sort | Kinosita, Yoshiaki |
collection | PubMed |
description | In many microorganisms helical structures are important for motility, e.g., bacterial flagella and kink propagation in Spiroplasma eriocheiris. Motile archaea also form a helical-shaped filament called the ‘archaellum’ that is functionally equivalent to the bacterial flagellum, but structurally resembles type IV pili. The archaellum motor consists of 6–8 proteins called fla accessory genes, and the filament assembly is driven by ATP hydrolysis at catalytic sites in FlaI. Remarkably, previous research using a dark-field microscopy showed that right-handed filaments propelled archaeal cells forwards or backwards by clockwise or counterclockwise rotation, respectively. However, the shape and rotational rate of the archaellum during swimming remained unclear, due to the low signal and lack of temporal resolution. Additionally, the structure and the motor properties of the archaellum and bacterial flagellum have not been precisely determined during swimming because they move freely in three-dimensional space. Recently, we developed an advanced method called “cross-kymography analysis”, which enables us to be a long-term observation and simultaneously quantify the function and morphology of helical structures using a total internal reflection fluorescence microscope. In this review, we introduce the basic idea of this analysis, and summarize the latest information in structural and functional characterization of the archaellum motor. |
format | Online Article Text |
id | pubmed-6018435 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | The Biophysical Society of Japan (BSJ) |
record_format | MEDLINE/PubMed |
spelling | pubmed-60184352018-06-28 Cross-kymography analysis to simultaneously quantify the function and morphology of the archaellum Kinosita, Yoshiaki Nishizaka, Takayuki Biophys Physicobiol Review Article In many microorganisms helical structures are important for motility, e.g., bacterial flagella and kink propagation in Spiroplasma eriocheiris. Motile archaea also form a helical-shaped filament called the ‘archaellum’ that is functionally equivalent to the bacterial flagellum, but structurally resembles type IV pili. The archaellum motor consists of 6–8 proteins called fla accessory genes, and the filament assembly is driven by ATP hydrolysis at catalytic sites in FlaI. Remarkably, previous research using a dark-field microscopy showed that right-handed filaments propelled archaeal cells forwards or backwards by clockwise or counterclockwise rotation, respectively. However, the shape and rotational rate of the archaellum during swimming remained unclear, due to the low signal and lack of temporal resolution. Additionally, the structure and the motor properties of the archaellum and bacterial flagellum have not been precisely determined during swimming because they move freely in three-dimensional space. Recently, we developed an advanced method called “cross-kymography analysis”, which enables us to be a long-term observation and simultaneously quantify the function and morphology of helical structures using a total internal reflection fluorescence microscope. In this review, we introduce the basic idea of this analysis, and summarize the latest information in structural and functional characterization of the archaellum motor. The Biophysical Society of Japan (BSJ) 2018-05-12 /pmc/articles/PMC6018435/ /pubmed/29955563 http://dx.doi.org/10.2142/biophysico.15.0_121 Text en 2018 © The Biophysical Society of Japan This article is licensed under the Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. To view a copy of this license, visit https://creativecommons.org/licenses/by-nc-sa/4.0/. |
spellingShingle | Review Article Kinosita, Yoshiaki Nishizaka, Takayuki Cross-kymography analysis to simultaneously quantify the function and morphology of the archaellum |
title | Cross-kymography analysis to simultaneously quantify the function and morphology of the archaellum |
title_full | Cross-kymography analysis to simultaneously quantify the function and morphology of the archaellum |
title_fullStr | Cross-kymography analysis to simultaneously quantify the function and morphology of the archaellum |
title_full_unstemmed | Cross-kymography analysis to simultaneously quantify the function and morphology of the archaellum |
title_short | Cross-kymography analysis to simultaneously quantify the function and morphology of the archaellum |
title_sort | cross-kymography analysis to simultaneously quantify the function and morphology of the archaellum |
topic | Review Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6018435/ https://www.ncbi.nlm.nih.gov/pubmed/29955563 http://dx.doi.org/10.2142/biophysico.15.0_121 |
work_keys_str_mv | AT kinositayoshiaki crosskymographyanalysistosimultaneouslyquantifythefunctionandmorphologyofthearchaellum AT nishizakatakayuki crosskymographyanalysistosimultaneouslyquantifythefunctionandmorphologyofthearchaellum |