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PMID: 11331302 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Insights into the dynamic properties of keratin intermediate filaments in living epithelial cells.

The Journal of cell biology ·Vol. 153 ·No. 3 ·2001-04-30 ·Pages 503-16

Yoon KH, Yoon M, Moir RD, Khuon S, Flitney FW, Goldman RD

Abstract

The properties of keratin intermediate filaments (IFs) have been studied after transfection with green fluorescent protein (GFP)-tagged K18 and/or K8 (type I/II IF proteins). GFP-K8 and -K18 become incorporated into tonofibrils, which are comprised of bundles of keratin IFs. These tonofibrils exhibit a remarkably wide range of motile and dynamic activities. Fluorescence recovery after photobleaching (FRAP) analyses show that they recover their fluorescence slowly with a recovery t(1/2) of approximately 100 min. The movements of bleach zones during recovery show that closely spaced tonofibrils (<1 microm apart) often move at different rates and in different directions. Individual tonofibrils frequently change their shapes, and in some cases these changes appear as propagated waveforms along their long axes. In addition, short fibrils, termed keratin squiggles, are seen at the cell periphery where they move mainly towards the cell center. The motile properties of keratin IFs are also compared with those of type III IFs (vimentin) in PtK2 cells. Intriguingly, the dynamic properties of keratin tonofibrils and squiggles are dramatically different from those of vimentin fibrils and squiggles within the same cytoplasmic regions. This suggests that there are different factors regulating the dynamic properties of different types of IFs within the same cytoplasmic regions.

MeSH Terms
Animals Antibodies/pharmacology Cells, Cultured Cytochalasin B/pharmacology Dyneins/immunology Energy Metabolism Epithelial Cells/physiology,radiation effects,ultrastructure Genes, Reporter Green Fluorescent Proteins Humans Intermediate Filaments/physiology,radiation effects,ultrastructure Keratins/physiology,ultrastructure Light Luminescent Proteins/radiation effects Microscopy, Fluorescence Movement/drug effects,physiology Nocodazole/pharmacology
Chemicals
Antibodies Luminescent Proteins Green Fluorescent Proteins Cytochalasin B Keratins Dyneins Nocodazole
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Yoon K H
Department of Cell and Molecular Biology, Northwestern University Medical School, Chicago, Illinois 60611.
Yoon M
Moir R D
Khuon S
Flitney F W
Goldman R D
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.153.3.503
Published
2001-04-30
Pages
503-16
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2190576
Subset
IM
Grants
NIDCR NIH HHS · 1 PO1 DE12328 · United States
External Links
PubMed source DOI: 10.1083/jcb.153.3.503 PMC Full Text View journal details
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