Self-assembly incompetence of synemin is related to the property of its head and rod domains.
The mechanisms regulating the intermediate filament (IF) protein assembly are complex and not yet fully understood. All vertebrate cytoplasmic IF proteins have a central alpha-helical rod domain flanked by variable head and tail domains. The IF protein synemin cannot homopolymerize to form filament networks; it needs an appropriate copolymerization partner. To elucidate the roles of the vimentin head domain, the TAAL motif in the 2A region, and the TYRKLLEGEE motif in the 2B region of the rod domain in synemin filament formation, we have prepared a series of synemin constructs by site-directed mutagenesis and chimeric synemins having the vimentin head domain. The assembly properties of synemin constructs were assessed by the immunofluorescence of transient transfection into cultured SW13 cells without endogenous IFs. Our data showed that the formation of a filamentous network required at least the vimentin-like head domain and both the 2A and 2B regions of the rod domain.
Khanamiryan
Luiza
L
UPMC Univ Paris 6, UMR 7079, Paris, France.
Li
Zhenlin
Z
Paulin
Denise
D
Xue
Zhigang
Z
D020816:Amino Acid Motifs / Q000502:physiology; D045744:Cell Line, Tumor; D006801:Humans; D007381:Intermediate Filament Proteins / Q000235:genetics / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D017354:Point Mutation; D017434:Protein Structure, Tertiary / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
D007381:Intermediate Filament Proteins; D011993:Recombinant Fusion Proteins; C430456:desmuslin