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Bellack, Annett ; Näther-Schindler, Daniela J. ; Schopf, Simone ; Rachel, Reinhard ; Wirth, Reinhard

Pyrococcus furiosus flagella: biochemical and transcriptional analyses identify the newly detected flaB0 gene to encode the major flagellin

Bellack, Annett, Näther-Schindler, Daniela J., Schopf, Simone, Rachel, Reinhard and Wirth, Reinhard (2014) Pyrococcus furiosus flagella: biochemical and transcriptional analyses identify the newly detected flaB0 gene to encode the major flagellin. Frontiers in Microbiology.

Date of publication of this fulltext: 26 Nov 2014 08:50
Article
DOI to cite this document: 10.5283/epub.31004


Abstract

We have described previously that the flagella of the Euryarchaeon Pyrococcus furiosus are multifunctional cell appendages used for swimming, adhesion to surfaces and formation of cell-cell connections. Here, we characterize these organelles with respect to their biochemistry and transcription. Flagella were purified by shearing from cells followed by CsCl-gradient centrifugation and were found ...

We have described previously that the flagella of the Euryarchaeon Pyrococcus furiosus are multifunctional cell appendages used for swimming, adhesion to surfaces and formation of cell-cell connections. Here, we characterize these organelles with respect to their biochemistry and transcription. Flagella were purified by shearing from cells followed by CsCl-gradient centrifugation and were found to consist mainly of a ca. 30 kDa glycoprotein. Polymerization studies of denatured flagella resulted in an ATP-independent formation of flagella-like filaments. The N-terminal sequence of the main flagellin was determined by Edman degradation, but none of the genes in the complete genome code for a protein with that N-terminus. Therefore, we resequenced the respective region of the genome, thereby discovering that the published genome sequence is not correct. A total of 771 bp are missing in the data base, resulting in the correction of the previously unusual N-terminal sequence of flagellin FlaB1 and in the identification of a third flagellin. To keep in line with the earlier nomenclature we call this flaB0. Very interestingly, the previously not identified flaB0 codes for the major flagellin. Transcriptional analyses of the revised flagellar operon identified various different cotranscripts encoding only a single protein in case of FlaB0 and FlaJ or up to five proteins (FlaB0-FlaD). Analysing the RNA of cells from different growth phases, we found that the length and number of detected cotranscript increased over time suggesting that the flagellar operon is transcribed mostly in late exponential and stationary growth phase.



Involved Institutions


Details

Item typeArticle
Journal or Publication TitleFrontiers in Microbiology
Publisher:FRONTIERS RESEARCH FOUNDATION
Place of Publication:LAUSANNE
Date24 November 2014
InstitutionsBiology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Mikrobiologie (Archaeenzentrum)
Identification Number
ValueType
10.3389/fmicb.2014.00695DOI
KeywordsMETHANOCOCCUS-MARIPALUDIS; ARCHAEAL FLAGELLUM; MOTILITY STRUCTURE; SURFACES; VOLTAE; GENOME; archaeal flagella; Pyrococcus furiosus; Fla proteins; major flagellin; FlaB0; transcriptional analyses
Dewey Decimal Classification500 Science > 570 Life sciences
StatusPublished
RefereedYes, this version has been refereed
Created at the University of RegensburgYes
URN of the UB Regensburgurn:nbn:de:bvb:355-epub-310040
Item ID31004

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