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Insights into subunit interactions in the S ulfolobus acidocaldarius archaellum cytoplasmic complex
Author(s) -
Banerjee Ankan,
Neiner Tomasz,
Tripp Patrick,
Albers SonjaVerena
Publication year - 2013
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/febs.12534
Subject(s) - sulfolobus acidocaldarius , cytoplasm , flagellum , protein subunit , s layer , chemistry , microscale thermophoresis , biophysics , biology , stereochemistry , microbiology and biotechnology , biochemistry , archaea , gene
Archaella are the archaeal motility structure that is the functional pendant of the bacterial flagellum but is assembled by a mechanism similar to that for type IV pili. Recently, it was shown by Banerjee et al . that F la X , a crenarchaeal archaellum subunit from Sulfolobus acidocaldarius , forms a ring‐like oligomer, and it was proposed that this ring may act as a static platform for torque generation in archaellum rotation [Banerjee A et al . (2012) J Biol Chem 287 , 43322–43330]. Moreover, the hexameric crystal structure of F la I was solved, and its dual function in the assembly and the rotation of the archaellum was demonstrated [Reindl S et al . (2013) Mol Cell 49 , 1069–1082]. In this study, we show by biochemical and biophysical techniques that F la X from S. acidocaldarius acts as a cytoplasmic scaffold in archaellum assembly, as it interacts with F la I as well as with the rec A family protein F la H , the only cytoplasmic components of the archaellum. Interaction studies using various truncated versions of F la I demonstrated that its N‐ and C‐termini interact with F la X . Moreover, using microscale thermophoresis, we show that F la I , F la X and F la H interact with high affinities in the nanomolar range. Therefore, we propose that these three proteins form the cytoplasmic motor complex of the archaellum. Structured digital abstractFlaH and FlaI bind by mst ( View interaction )FlaXc physically interacts with FlaI by pull down ( 1 , 2 )FlaI and FlaXc bind by mst ( 1 , 2 )FlaI binds to FlaXc by pull down ( View interaction )