|
|
|
|
|
|
|
|
|||||
| Home Help Feedback Subscriptions Archive Search Table of Contents | |||||
Journal of Cell Science, Vol 105, Issue 2 433-444, Copyright © 1993 by Company of Biologists
JOURNAL ARTICLES |
CF Chou, CL Riopel, LS Rott and MB Omary
Palo Alto Veterans Administration Medical Center, CA.
We studied the solubility of keratin polypeptides 8 and 18 (K8/18), which are the predominant intermediate filaments in the human colonic epithelial cell line HT29. We find that asynchronously growing cells (G0/G1 stage of the cell cycle) have a substantial pool of soluble keratin that constitutes approx. 5% of total cellular keratin. This soluble keratin pool was observed after immunoprecipitation of K8/18 from the cytosolic fraction of cells disrupted using three detergent-free methods. Several other cell lines showed a similar significant soluble cytosolic K8/18 pool. Arrest of HT29 cells in G2/M stage of the cell cycle was associated with a concurrent increase in keratin solubility. Comparison of K8/18 obtained from the soluble cytosolic fraction and the insoluble high-speed pellet fraction showed similar levels of phosphorylation and glycosylation and similar tryptic radiolabeled phospho- and glycopeptide patterns. Soluble K8/18 can form characteristic 10 nm filaments in vitro as determined by electron microscopy. Cross-linking of soluble K8/18 followed by immunoprecipitation resulted in dimeric and tetrameric forms, based on migration in SDS-polyacrylamide gels. In addition, cross-linked and native soluble K8/18 showed similar migration on nondenaturing gels and similar sedimentation after sucrose density gradient centrifugation. Our results indicate that simple epithelial keratins are appreciably more soluble than previously recognized. The soluble keratin form is assembly competent and appears to be primarily tetrameric. Although K8/18 solubility was found to increase during mitotic arrest, glycosylation and phosphorylation did not play an obvious role in generating the soluble fraction, suggesting an alternate mechanism for keratin solubility.
This article has been cited by other articles:
|
|
 |
|
  S. Kim and P. A. Coulombe Intermediate filament scaffolds fulfill mechanical, organizational, and signaling functions in the cytoplasm Genes & Dev., July 1, 2007; 21(13): 1581 - 1597. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Sen, D. Agresti, and E. R. Mackow Hyperphosphorylation of the Rotavirus NSP5 Protein Is Independent of Serine 67 or NSP2, and the Intrinsic Insolubility of NSP5 Is Regulated by Cellular Phosphatases J. Virol., February 15, 2006; 80(4): 1807 - 1816. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. K. Scott, C. Atsriku, P. Kaminker, J. Held, B. Gibson, M. A. Baldwin, and C. C. Benz Vitamin K3 (Menadione)-Induced Oncosis Associated with Keratin 8 Phosphorylation and Histone H3 Arylation Mol. Pharmacol., September 1, 2005; 68(3): 606 - 615. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. M. Bernot, C.-H. Lee, and P. A. Coulombe A small surface hydrophobic stripe in the coiled-coil domain of type I keratins mediates tetramer stability J. Cell Biol., March 14, 2005; 168(6): 965 - 974. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L.-H. Gu and P. A. Coulombe Defining the Properties of the Nonhelical Tail Domain in Type II Keratin 5: Insight from a Bullous Disease-causing Mutation Mol. Biol. Cell, March 1, 2005; 16(3): 1427 - 1438. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. Zhong, Q. Zhou, D. M. Toivola, G.-Z. Tao, E. Z. Resurreccion, and M. B. Omary Organ-specific stress induces mouse pancreatic keratin overexpression in association with NF-{kappa}B activation J. Cell Sci., May 1, 2004; 117(9): 1709 - 1719. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. S. Werner, R. Windoffer, P. Strnad, C. Grund, R. E. Leube, and T. M. Magin Epidermolysis Bullosa Simplex-Type Mutations Alter the Dynamics of the Keratin Cytoskeleton and Reveal a Contribution of Actin to the Transport of Keratin Subunits Mol. Biol. Cell, March 1, 2004; 15(3): 990 - 1002. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Q. Wang, H. Griffin, S. Southern, D. Jackson, A. Martin, P. McIntosh, C. Davy, P. J. Masterson, P. A. Walker, P. Laskey, et al. Functional Analysis of the Human Papillomavirus Type 16 E1{wedge}E4 Protein Provides a Mechanism for In Vivo and In Vitro Keratin Filament Reorganization J. Virol., January 15, 2004; 78(2): 821 - 833. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G.-Z. Tao, D. M. Toivola, B. Zhong, S. A. Michie, E. Z. Resurreccion, Y. Tamai, M. M. Taketo, and M. B. Omary Keratin-8 null mice have different gallbladder and liver susceptibility to lithogenic diet-induced injury J. Cell Sci., November 15, 2003; 116(22): 4629 - 4638. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Q. Zhou, D. M. Toivola, N. Feng, H. B. Greenberg, W. W. Franke, and M. B. Omary Keratin 20 Helps Maintain Intermediate Filament Organization in Intestinal Epithelia Mol. Biol. Cell, July 1, 2003; 14(7): 2959 - 2971. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Liovic, M. M. Mogensen, A. R. Prescott, and E. B. Lane Observation of keratin particles showing fast bidirectional movement colocalized with microtubules J. Cell Sci., April 15, 2003; 116(8): 1417 - 1427. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N.-O. Ku, S. Azhar, and M. B. Omary Keratin 8 Phosphorylation by p38 Kinase Regulates Cellular Keratin Filament Reorganization. MODULATION BY A KERATIN 1-LIKE DISEASE-CAUSING MUTATION J. Biol. Chem., March 22, 2002; 277(13): 10775 - 10782. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Strnad, R. Windoffer, and R. E. Leube Induction of rapid and reversible cytokeratin filament network remodeling by inhibition of tyrosine phosphatases J. Cell Sci., January 11, 2002; 115(21): 4133 - 4148. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. H. Yoon, M. Yoon, R. D. Moir, S. Khuon, F. W. Flitney, and R. D. Goldman Insights into the Dynamic Properties of Keratin Intermediate Filaments in Living Epithelial Cells J. Cell Biol., April 24, 2001; 153(3): 503 - 516. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Bertelli, M. Regoli, F. Gambelli, M. Lucattelli, G. Lungarella, and A. Bastianini GFAP Is Expressed as a Major Soluble Pool Associated with Glucagon Secretory Granules in A-cells of Mouse Pancreas J. Histochem. Cytochem., September 1, 2000; 48(9): 1233 - 1242. [Abstract] [Full Text]
|
|
|
|
|
|
N.-O. Ku and M. B. Omary Keratins Turn Over by Ubiquitination in a Phosphorylation-modulated Fashion J. Cell Biol., May 1, 2000; 149(3): 547 - 552. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R Windoffer and R. Leube Detection of cytokeratin dynamics by time-lapse fluorescence microscopy in living cells J. Cell Sci., January 12, 1999; 112(24): 4521 - 4534. [Abstract] [PDF]
|
|
|
|
|
|
J. Fradette, L. Germain, P. Seshaiah, and P. A. Coulombe The Type I Keratin 19 Possesses Distinct and Context-dependent Assembly Properties J. Biol. Chem., December 25, 1998; 273(52): 35176 - 35184. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. A. Spencer, A. S. Coutts, S. K. Samuel, L. C. Murphy, and J. R. Davie Estrogen Regulates the Association of Intermediate Filament Proteins with Nuclear DNA in Human Breast Cancer Cells J. Biol. Chem., October 30, 1998; 273(44): 29093 - 29097. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Wawersik, R. D. Paladini, E. Noensie, and P. A. Coulombe A Proline Residue in the alpha -Helical Rod Domain of Type I Keratin 16 Destabilizes Keratin Heterotetramers J. Biol. Chem., December 19, 1997; 272(51): 32557 - 32565. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N.-O. Ku and M. B. Omary Phosphorylation of Human Keratin 8in Vivo at Conserved Head Domain Serine 23and at Epidermal Growth Factor-stimulated Tail Domain Serine 431 J. Biol. Chem., March 14, 1997; 272(11): 7556 - 7564. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Paramio, M. Casanova, A Alonso, and J. Jorcano Keratin intermediate filament dynamics in cell heterokaryons reveals diverse behaviour of different keratins J. Cell Sci., January 5, 1997; 110(9): 1099 - 1111. [Abstract] [PDF]
|
|
|
|
|
|
R Pankov, I Simcha, M Zoller, R. Oshima, and A Ben-Ze'ev Contrasting effects of K8 and K18 on stabilizing K19 expression, cell motility and tumorigenicity in the BSp73 adenocarcinoma J. Cell Sci., January 4, 1997; 110(8): 965 - 974. [Abstract] [PDF]
|
|
|
|
|
|
N.-O. Ku and M. B. Omary Identification and Mutational Analysis of the Glycosylation Sites of Human Keratin 18 J. Biol. Chem., May 19, 1995; 270(20): 11820 - 11827. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Liao, L. A. Lowthert, N. Ghori, and M. B. Omary The 70-kDa Heat Shock Proteins Associate with Glandular Intermediate Filaments in an ATP-dependent Manner J. Biol. Chem., January 13, 1995; 270(2): 915 - 922. [Abstract] [Full Text] [PDF]
|
|
