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J. Biol. Chem., Vol. 279, Issue 9, 8181-8189, February 27, 2004

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HLS5, a Novel RBCC (Ring Finger, B Box, Coiled-coil) Family Member Isolated from a Hemopoietic Lineage Switch, Is a Candidate Tumor Suppressor^*

Jean-Philippe Lalonde, Raelene Lim¶||, Evan Ingley, Peta A. Tilbrook, Martin J. Thompson, Ross McCulloch, Jennifer G Beaumont, Carol Wicking**, Helen J. Eyre, Grant R. Sutherland, Kathy Howe, Ellen Solomon¶¶, James H. Williams, and S. Peter Klinken||||

From the Laboratory for Cancer Medicine, Western Australian Institute for Medical Research, Royal Perth Hospital and the Center for Medical Research, The University of Western Australia, Perth, Western Australia 6000, Australia, ^¶School of Biomedical Sciences, Curtin University of Technology, Bentley, Western Australia 6102, Australia, ^**Institute for Molecular Bioscience, University of Queensland, Brisbane, Queensland 4072, Australia, Center for Medical Genetics, Department of Cytogenetics and Molecular Genetics, Women's Hospital, Adelaide, Southern Australia 5006, Australia, Lymphocyte Activation Laboratory, Cancer Research UK, 44 Lincoln's Inn Fields, London WC2A 3PX, United Kingdom, and ^¶¶Division of Medical and Molecular Genetics, Guy's School of Medicine, King's School of Medicine, and St. Thomas' School of Medicine, London SE1 9RT, United Kingdom

Hemopoietic cells, apparently committed to one lineage, can be reprogrammed to display the phenotype of another lineage. The J2E erythroleukemic cell line has on rare occasions developed the features of monocytic cells. Subtractive hybridization was used in an attempt to identify genes that were up-regulated during this erythroid to myeloid transition. We report here on the isolation of hemopoietic lineage switch 5 (Hls5), a gene expressed by the monocytoid variant cells, but not the parental J2E cells. Hls5 is a novel member of the RBCC (Ring finger, B box, coiled-coil) family of genes, which includes Pml, Herf1, Tif-1, and Rfp. Hls5 was expressed in a wide range of adult tissues; however, at different stages during embryogenesis, Hls5 was detected in the branchial arches, spinal cord, dorsal root ganglia, limb buds, and brain. The protein was present in cytoplasmic granules and punctate nuclear bodies. Isolation of the human cDNA and genomic DNA revealed that the gene was located on chromosome 8p21, a region implicated in numerous leukemias and solid tumors. Enforced expression of Hls5 in HeLa cells inhibited cell growth, clonogenicity, and tumorigenicity. It is conceivable that HLS5 is one of the tumor suppressor genes thought to reside at the 8p21 locus.

Received for publication, June 25, 2003 , and in revised form, December 5, 2003.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) AF009514, AF009513, AAC17945 AF145374, AF494189, and AF492463.

^* This work was supported by the National Health and Medical Research Council of Australia (Grant 212070), the National Breast Cancer Foundation, the Cancer Foundation of Western Australia, the Medical Research Foundation of Royal Perth Hospital, and Cancer Research UK. 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.

Recipient of an M.D. and Ph.D. Ad Hoc scholarship awarded by the Stalker McEwan fund through the Medical Research Foundation of Royal Perth Hospital.

^|| Recipient of an Australian Postgraduate Award through Curtin University of Technology.

^|||| To whom correspondence should be addressed: Laboratory for Cancer Medicine, Level 6, Medical Research Foundation Bldg., Rear, 50 Murray St., Perth, Western Australia 6000, Australia. Tel.: 61-8-92240334; Fax: 61-8-92240322; E-mail: pklinken{at}waimr.uwa.edu.au.

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