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Matrix-mediated changes in the expression of HNF-4alpha isoforms and in DNA-binding activity of ARP-1 in primary cultures of rat hepatocytes.
Biochem Biophys Res Commun. 1999 Jun 16; 259(3):651-5.BB

Abstract

Recently, we have developed a culture system in which rat hepatocytes dedifferentiate and proliferate and after the addition of EHS-gel redifferentiate. During both developmental stages HNF-4alpha2 mRNA was more abundant than HNF-4alpha1 mRNA. However, Western blot analysis using COS-7 cell-expressed HNF-4alpha1 and HNF-4alpha2 proteins as standards revealed that (i) HNF-4alpha2 protein was not expressed in dedifferentiated hepatocytes and (ii) either HNF-4alpha2 protein or a highly phosphorylated HNF-4alpha1 protein was the dominating isoform in redifferentiated hepatocytes. The changes in HNF4-isoform expression could not be mimicked by DMSO, suggesting them to be matrix specific. Furthermore, DMSO was less efficient than EHS-gel in reinducing liver-specific gene expression. EHS-gel overlay also led to reduction of ARP-1 DNA binding activity, while overall ARP-1 protein levels did not change. These results suggest that EHS-matrix overlay regulates the expression of different HNF-4alpha isoforms on a posttranscriptional level while ARP-1 DNA binding activity is regulated by posttranslational mechanisms.

Authors+Show Affiliations

Department of Pathology, University of Pittsburgh, Pittsburgh, Pennsylvania, 15261, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

10364473

Citation

Runge, D, et al. "Matrix-mediated Changes in the Expression of HNF-4alpha Isoforms and in DNA-binding Activity of ARP-1 in Primary Cultures of Rat Hepatocytes." Biochemical and Biophysical Research Communications, vol. 259, no. 3, 1999, pp. 651-5.
Runge D, Runge DM, Daskalakis N, et al. Matrix-mediated changes in the expression of HNF-4alpha isoforms and in DNA-binding activity of ARP-1 in primary cultures of rat hepatocytes. Biochem Biophys Res Commun. 1999;259(3):651-5.
Runge, D., Runge, D. M., Daskalakis, N., Lubecki, K. A., Bowen, W. C., & Michalopoulos, G. K. (1999). Matrix-mediated changes in the expression of HNF-4alpha isoforms and in DNA-binding activity of ARP-1 in primary cultures of rat hepatocytes. Biochemical and Biophysical Research Communications, 259(3), 651-5.
Runge D, et al. Matrix-mediated Changes in the Expression of HNF-4alpha Isoforms and in DNA-binding Activity of ARP-1 in Primary Cultures of Rat Hepatocytes. Biochem Biophys Res Commun. 1999 Jun 16;259(3):651-5. PubMed PMID: 10364473.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Matrix-mediated changes in the expression of HNF-4alpha isoforms and in DNA-binding activity of ARP-1 in primary cultures of rat hepatocytes. AU - Runge,D, AU - Runge,D M, AU - Daskalakis,N, AU - Lubecki,K A, AU - Bowen,W C, AU - Michalopoulos,G K, PY - 1999/6/12/pubmed PY - 1999/6/12/medline PY - 1999/6/12/entrez SP - 651 EP - 5 JF - Biochemical and biophysical research communications JO - Biochem Biophys Res Commun VL - 259 IS - 3 N2 - Recently, we have developed a culture system in which rat hepatocytes dedifferentiate and proliferate and after the addition of EHS-gel redifferentiate. During both developmental stages HNF-4alpha2 mRNA was more abundant than HNF-4alpha1 mRNA. However, Western blot analysis using COS-7 cell-expressed HNF-4alpha1 and HNF-4alpha2 proteins as standards revealed that (i) HNF-4alpha2 protein was not expressed in dedifferentiated hepatocytes and (ii) either HNF-4alpha2 protein or a highly phosphorylated HNF-4alpha1 protein was the dominating isoform in redifferentiated hepatocytes. The changes in HNF4-isoform expression could not be mimicked by DMSO, suggesting them to be matrix specific. Furthermore, DMSO was less efficient than EHS-gel in reinducing liver-specific gene expression. EHS-gel overlay also led to reduction of ARP-1 DNA binding activity, while overall ARP-1 protein levels did not change. These results suggest that EHS-matrix overlay regulates the expression of different HNF-4alpha isoforms on a posttranscriptional level while ARP-1 DNA binding activity is regulated by posttranslational mechanisms. SN - 0006-291X UR - https://www.unboundmedicine.com/medline/citation/10364473/Matrix_mediated_changes_in_the_expression_of_HNF_4alpha_isoforms_and_in_DNA_binding_activity_of_ARP_1_in_primary_cultures_of_rat_hepatocytes_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0006-291X(99)90848-3 DB - PRIME DP - Unbound Medicine ER -