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J. Biol. Chem., Vol. 279, Issue 26, 27753-27763, June 25, 2004

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Cell Type-specific Expression of LINE-1 Open Reading Frames 1 and 2 in Fetal and Adult Human Tissues^*

Süleyman Ergün, Christian Buschmann¶||, Jochen Heukeshoven**, Kristin Dammann¶, Frank Schnieders¶, Heidrun Lauke, Fariba Chalajour, Nerbil Kilic, Wolf H. Strätling¶, and Gerald G. Schumann¶||

From the Institut für Anatomie, the ^¶Institut für Biochemie und Molekularbiologie, and the Klinik und Poliklinik der Inneren Medizin, Universitätsklinikum Hamburg-Eppendorf, Martinistrasse 52, Hamburg D-20246, ^||Section Pr2/Retroelements, Paul-Ehrlich-Institut, Paul-Ehrlich-Strasse 51-59, Langen D-63225, and the ^**Heinrich-Pette-Institut für Experimentelle Virologie und Immunologie an der Universität Hamburg, Martinistrasse 52, Hamburg D-20206, Germany

The LINE-1 (L1) family of non-long terminal repeat retrotransposons is a major force shaping mammalian genomes, and its members can alter the genome in many ways. Mutational analyses have shown that coexpression of functional proteins encoded by the two L1-specific open reading frames, ORF1 and ORF2, is an essential prerequisite for the propagation of L1 elements in the genome. However, all efforts to identify ORF2-encoded proteins have failed so far. Here, applying a novel antibody we report the presence of proteins encoded by ORF2 in a subset of cellular components of human male gonads. Immunohistochemical analyses revealed coexpression of ORF1 and ORF2 in prespermatogonia of fetal testis, in germ cells of adult testis, and in distinct somatic cell types, such as Leydig, Sertoli, and vascular endothelial cells. Coexpression of both proteins in male germ cells is necessary for the observed genomic expansion of the number of L1 elements. Peptide mass fingerprinting analysis of a 130-kDa polypeptide isolated from cultured human dermal microvascular endothelial cells led to the identification of ORF2-encoded peptides. An isolated 45-kDa polypeptide was shown to derive from nonfunctional copies of ORF2 coding regions. The presence of both ORF1- and ORF2-encoded proteins in vascular endothelial cells and its apparent association with certain stages of differentiation and maturation of blood vessels may have functional relevance for vasculogenesis and/or angiogenesis.

Received for publication, November 30, 2003 , and in revised form, March 30, 2004.

^* This work was supported by Grants Schu 1014/2-1 and 1014/2-4 (to G. G. S.) and SFB545/B2 (to W. H. S.) from the Deutsche Forschungsgemeinschaft and Grant Az.10.01.1.104 from the Fritz-Thyssen-Stiftung (to G. G. S.). 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.

These authors contributed equally to this work.

To whom correspondence should be addressed: Section Pr2/Retro-elements, Paul-Ehrlich-Institut, Paul-Ehrlich-Strasse 51-59, Langen D-63225, Germany. Tel.: 49-6103-773-105; Fax: 49-6103-771-265; E-mail: schgr{at}pei.de.

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