Flagellar structure and hyperthermophily: analysis of a single flagellin gene and its product in Aquifex pyrophilus.
The polytrichously inserted flagella of Aquifex pyrophilus, a marine hyperthermophilic bacterium growing at 85 degrees C, were isolated and purified. Electron micrographs of the 19-nm-diameter flagellar filaments show prominent helical arrays of subunits. The primary structure of these 54-kDa flagellin monomers determining the helical shape and heat stability of filaments was of particular interest. The genomic region encoding the flagellin subunit (flaA gene) and an upstream open reading frame (orf1) were cloned and sequenced. The 1,503-bp flaA and 696-bp orf1 are preceded by separate sigma 28-like promoters and ribosome-binding motifs and succeeded by palindromic transcription terminators. Both genes are actively transcribed, but the nature and function of the orf1-encoded 231-residue polypeptide remain unknown. The deduced primary structure of the 501-amino-acid flagellin encoded by flaA consists of conserved N- and C-terminal regions and a variable 246-residue central domain. In comparison to mesophilic flagellins, the thermostable A. pyrophilus flagellin is characterized by increases in aromatic residues and prolines as well as by a 7.9% +/- 3.2% increase in all hydrophobic residues that is balanced by a respective decrease in hydrophilic residues. This composition is thought to form more compact flagellin monomers and stable interface contacts between neighboring subunits in the polymer.
Behammer
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W
Lehrstuhl für Genetik, University of Regensburg, Germany.
Shao
Z
Z
Mages
W
W
Rachel
R
R
Stetter
K O
KO
Schmitt
R
R
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001483:Base Sequence; D003001:Cloning, Molecular; D005407:Flagella / Q000737:chemistry / Q000648:ultrastructure*; D005408:Flagellin / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000648:ultrastructure; D018628:Gene Dosage; D005798:Genes, Bacterial / Q000235:genetics*; D006088:Gram-Negative Aerobic Bacteria / Q000737:chemistry* / Q000235:genetics; D006358:Hot Temperature; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D012329:RNA, Bacterial / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D015183:Restriction Mapping; D017421:Sequence Analysis; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D000596:Amino Acids; D012329:RNA, Bacterial; D012333:RNA, Messenger; D005408:Flagellin; C067138:flaA protein, bacteria