Microscopic, genetic, and biochemical characterization of non-flagellar swimming motility in marine cyanobacteria
- Author(s): McCarren, Jay William
- et al.
The mechanism of motility in marine Synechococcus which swim without any apparent extracellular appendages, remains a mystery 20 years after its discovery. A multifaceted investigation including direct microscopic visualization, genetic analyses, and biochemical approaches was carried out in order to better understand the physiology of this globally important primary producer. Ultrastructural analyses provided a detailed view of the cell envelope layers and aided in the identification of a structure important for motility. Electron microscope tomographic reconstructions revealed the even distribution of SwmA, a protein required for motility, across the cell surface. Various cryo-fixation techniques were required for the preservation and visualization of a para-crystalline S-layer formed by this protein. As complete genomic sequence information failed to identify genes involved in motility, a transposon mutagenesis technique was developed to identify components of the motility apparatus. Utilizing this genetic tool, 17 independent transposon insertions that abolish motility were localized to clusters in three separate chromosomal regions. Included within these clusters are several multicomponent transport systems, as well as a number of glycosyltranferases. One cluster is characterized by DNA with an exceptionally low % G+C content relative to the genome average. Additionally, inter-genome comparisons reveal the absence of this stretch of DNA in two non- motile strains of Synechococcus suggesting acquisition of this genetic information by horizontal gene transfer. Contained within this region of low % G+C content is an extremely large gene called SwmB, which is required for motility in these cells. The sequence of SwmB is highly repetitive, with 4 domains of tandem repeats comprising over 60% of the protein. Analyses confirm that this gene is indeed translated into a megadalton-size protein, which is localized on the cell surface. Cellular localization of the two motility proteins SwmA and SwmB revealed that all motility mutants in culture have a defect in the localization of either SwmA or SwmB and in some instances both of these proteins. Additionally, two outer membrane polypeptides of 70 kDa and 80 kDa are absent in some of these mutants, suggestive of a role in motility