Chaperonin filaments: the archaeal cytoskeleton?
Chaperonins are high molecular mass double-ring structures composed of 60-kDa protein subunits. In the hyperthermophilic archaeon Sulfolobus shibatae the two chaperonin proteins represent approximately 4% of its total protein and have a combined intracellular concentration of >30 mg/ml. At concentrations >/= 0.5 mg/ml purified chaperonins form filaments in the presence of Mg2+ and nucleotides. Filament formation requires nucleotide binding (not hydrolysis), and occurs at physiological temperatures in biologically relevant buffers, including a buffer made from cell extracts. These observations suggest that chaperonin filaments may exist in vivo and the estimated 4600 chaperonins per cell suggest that such filaments could form an extensive cytostructure. We observed filamentous structures in unfixed, uranyl-acetate-stained S. shibatae cells, which resemble the chaperonin filaments in size and appearance. ImmunoGold (Janssen) labeling using chaperonin antibodies indicated that many chaperonins are associated with insoluble cellular structures and these structures appear to be filamentous in some areas, although they could not be uranyl-acetate-stained. The existence of chaperonin filaments in vivo suggests a mechanism whereby their protein-folding activities can be regulated. More generally, the filaments themselves may play a cytoskeletal role in Archaea.
Trent
J D
JD
Center for Mechanistic Biology and Biotechnology, Argonne National Laboratory, 9700 South Cass Avenue, Argonne, IL 60439, USA. Trent@ANLCMB.bim.anl.gov
Kagawa
H K
HK
Yaoi
T
T
Olle
E
E
Zaluzec
N J
NJ
D008841:Actin Cytoskeleton / Q000648:ultrastructure; D000244:Adenosine Diphosphate / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000494:pharmacology; D000266:Adenylyl Imidodiphosphate / Q000494:pharmacology; D018833:Chaperonins / Q000187:drug effects / Q000302:isolation & purification / Q000648:ultrastructure*; D003599:Cytoskeleton / Q000648:ultrastructure*; D008274:Magnesium / Q000494:pharmacology; D008854:Microscopy, Electron; D016253:Microscopy, Immunoelectron; D016944:Sulfolobus / Q000648:ultrastructure*
D000266:Adenylyl Imidodiphosphate; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D018833:Chaperonins; D008274:Magnesium