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J. Biol. Chem., Vol. 279, Issue 9, 8063-8069, February 27, 2004
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¶¶
From the
Institute of Zoology, the Institute of Molecular Genetics, the **Institute of Physiological Chemistry, and the Department of Anatomy, Johannes Gutenberg-University, D-55099 Mainz, Germany, the ¶Institute of Evolution, University of Haifa, Mt. Carmel, Haifa 31905, Israel, and ||Cell Biology and Biophysics, European Molecular Biology Laboratory, D-69117 Heidelberg, Germany
Cytoglobin is a recently discovered vertebrate globin distantly related to myoglobin, and its function is unknown. Here we present the first detailed analysis of the distribution and expression of cytoglobin. Northern and Western blotting experiments show the presence of cytoglobin mRNA and protein in a broad range of tissues. Quantitative PCR demonstrates an up-regulation of cytoglobin mRNA levels in rat heart and liver under hypoxic conditions (22 and 44 h of 9% oxygen). Immunofluorescence studies with three antibodies directed against different epitopes of the protein consistently show cytoglobin in connective tissue fibroblasts as well as in hepatic stellate cells. Cytoglobin is also present in chondroblasts and osteoblasts and shows a decreased level of expression upon differentiation to chondrocytes and osteocytes. Cytoglobin is located in the cytoplasm of these cell types. Evidence against an exclusively nuclear localization of cytoglobin, as recently proposed, is also provided by transfection assays with green fluorescent protein fusion constructs, which demonstrates the absence of an active nuclear import. The differential expression of cytoglobin argues against a general respiratory function of this molecule, but rather indicates a connective tissue-specific function. We hypothesize that cytoglobin may be involved in collagen synthesis. Cytoglobin expression was also observed in some neuronal subpopulations of the central and the peripheral nervous systems. Surprisingly, cytoglobin is localized in both the cytoplasm and nucleus of neurons, indicating a possible additional role of this protein in neuronal tissues.
Received for publication, September 23, 2003 , and in revised form, December 2, 2003.
* This work was supported in part by the Deutsche Forschungsgemeinschaft (Grants Bu956/5 and Ha2103/3) and the European Union (QLRT-2001-01548). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
The on-line version of this article (available at http://www.jbc.org) contains Supplementary Data.
¶¶ To whom correspondence should be addressed: Institute of Zoology, Molecular Animal Physiology, University of Mainz, Müllerweg 6, D-55099 Mainz, Germany. Tel.: 49-6131-39-24477; Fax: 49-6131-39-24652; E-mail: burmeste{at}uni-mainz.de.
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