The function of intermediate filaments in cell shape and cytoskeletal integrity.
This study describes the development and use of a specific method for disassembling intermediate filament (IF) networks in living cells. It takes advantage of the disruptive effects of mimetic peptides derived from the amino acid sequence of the helix initiation 1A domain of IF protein chains. The results demonstrate that at 1:1 molar ratios, these peptides disassemble vimentin IF into small oligomeric complexes and monomers within 30 min at room temperature in vitro. Upon microinjection into cultured fibroblasts, these same peptides induce the rapid disassembly of IF networks. The disassembly process is accompanied by a dramatic alteration in cell shape and the destabilization of microtubule and actin-stress fiber networks. These changes in cell shape and IF assembly states are reversible. The results are discussed with respect to the roles of IF in cell shape and the maintenance of the integrity and mechanical properties of the cytoplasm, as well as the stability of the other major cytoskeletal systems.
Goldman
R D
RD
Department of Cell and Molecular Biology, Northwestern University Medical School, Chicago, Illinois 60611, USA.
Khuon
S
S
Chou
Y H
YH
Opal
P
P
Steinert
P M
PM
D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D048429:Cell Size / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D003078:Colchicine / Q000494:pharmacology; D006224:Cricetinae; D003598:Cytoskeletal Proteins / Q000096:biosynthesis; D003599:Cytoskeleton / Q000378:metabolism*; D005347:Fibroblasts; D007382:Intermediate Filaments / Q000502:physiology*; D007668:Kidney; D051379:Mice; D008845:Microinjections; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000494:pharmacology*; D011108:Polymers; D014746:Vimentin / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology
D003598:Cytoskeletal Proteins; D010446:Peptide Fragments; D011108:Polymers; D014746:Vimentin; D003078:Colchicine