LIM-domain protein cysteine- and glycine-rich protein 2 (CRP2) is a novel marker of hepatic stellate cells and binding partner of the protein inhibitor of activated STAT1.
Activation of hepatic stellate cells is considered to be the main step in the development of liver fibrosis, which is characterized by the transition of quiescent vitamin-A-rich cells to proliferative, fibrogenic and contractile myofibroblasts. The identification of regulatory genes during early cell activation and transdifferentiation is essential to extend our knowledge of hepatic fibrogenesis. In liver, the gene CSRP2 is exclusively expressed by stellate cells, whereas no transcripts are detectable in hepatocytes, sinusoidal endothelial cells or Kupffer cells. The early activation of stellate cells induced by platelet-derived growth factor is accompanied by an enhanced expression of CSRP2. During later stages of transdifferentiation, the expression of CSRP2 in these cells is suppressed in vitro and in vivo. The CSRP2-encoded cysteine- and glycine-rich double-LIM-domain protein (CRP)2 is proposed to function as a molecular adapter, arranging two or more as yet unidentified protein constituents into a macromolecular complex. To identify these proteins and assign a cellular function to CRP2, a human cDNA library was screened with full-length CRP2 as bait in a yeast two-hybrid screen. The protein inhibitor of activated STAT1 ('PIAS1') was shown to associate selectively with the C-terminal LIM domain of CRP2. Physical interaction of both proteins in the cellular environment was confirmed by co-localization experiments with confocal laser scanning microscopy and co-immunoprecipitation analysis. These results establish CRP2 as a potential new factor in the JAK/STAT-signalling pathway and suggest that the suppression of CSRP2 might be a prerequisite for the myofibroblastic transition of hepatic stellate cells.
Weiskirchen
R
R
Institute of Clinical Chemistry and Pathobiochemistry, RWTH-University Hospital, Pauwelsstrasse 30, D-52074 Aachen, Germany. rweiskirchen@post.klinikum.rwth-aachen.de
Moser
M
M
Weiskirchen
S
S
Erdel
M
M
Dahmen
S
S
Buettner
R
R
Gressner
A M
AM
D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D015415:Biomarkers; D002478:Cells, Cultured; D002874:Chromosome Mapping; D004622:Embryo, Mammalian / Q000378:metabolism; D022781:Hepatocytes / Q000378:metabolism; D006801:Humans; D017403:In Situ Hybridization; D060588:LIM Domain Proteins; D008099:Liver / Q000166:cytology* / Q000378:metabolism; D008103:Liver Cirrhosis / Q000473:pathology; D008297:Male; D051379:Mice; D018613:Microscopy, Confocal; D009687:Nuclear Proteins; D011485:Protein Binding; D050828:Protein Inhibitors of Activated STAT; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction / Q000502:physiology; D020798:Two-Hybrid System Techniques; D016335:Zinc Fingers / Q000235:genetics
D048868:Adaptor Proteins, Signal Transducing; D015415:Biomarkers; C486176:CRIP2 protein, human; D060588:LIM Domain Proteins; D009687:Nuclear Proteins; C494211:Pias1 protein, mouse; D050828:Protein Inhibitors of Activated STAT; D011506:Proteins; D016271:Proto-Oncogene Proteins c-myc; C074370:cysteine-rich protein, mammalian