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Deletion analysis of chromosome 13q14.3 and characterisation of an alternative splice form of LEU1 in B cell chronic lymphocytic leukemia

Leukemia volume 16, pages 1267–1275 (2002)Cite this article

Abstract

Heterozygous and homozygous deletions of chromosome 13q14.3 are found in 50% of patients with B cell CLL, suggesting the presence of one or more tumour suppressor genes within the deleted region. To identify candidate genes from the region, we constructed a map of 13q14.3 using a combination of genomic and cDNA library screening. The incidence of deletions in CLL patients was 51.5% encompassing a 265 kb region of minimal deletion (RMD) telomeric to markers D13S319. Two CpG islands were identified within the RMD, the telomeric of which is fully methylated whilst the more centromeric is unmethylated. A novel transcript was identified within the RMD that represents an alternative splice version of Leu1. The nine exons of this transcript span a genomic of 436 kb with exon 1 of Leu1 being the common first exon. The remaining exons were shown to be more frequently deleted than Leu1 itself. All splice forms of this transcript were detectable by RT-PCR but Leu1 detected the most abundant message on Northern blotting. Sequence analysis failed to reveal inactivating mutations in patients with heterozygous deletion of 13q14.3, although a polymorphic T to A variant was identified within exon 1 of Leu1 in leukemic and normal controls. As no mutations have been detected for Leu1 or any other transcript so far described, we cannot exclude the existence of control elements within the RMD that may regulate expression of genes lying in this region.

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Acknowledgements

We would like to thank Dr Atul Mehta, MD, Professor HG Prentice, K Ganeshaguru, PhD and Steve Hart, PhD (Department of Hematology, Royal Free and University College School of Medicine) for assisting in the collection of material from CLL patients; Professor P Sfikakis (University of Athens, Laikon Hospital, Athens Greece) for financial support to Dr Panayiotidis and Dr Kotsi; Ann Walker, PhD (Department of Medicine, Royal Free and University College School of Medicine) for critically reviewing the manuscript and providing DNA from normal controls, and Andrew Dunham, PhD (The Sanger Centre, Hinxton, Cambs, UK) for invaluable help with the sequencing of BAC clones and gene analysis of chromosome 13. Research grant support: Kay Kendall Leukaemia Fund.

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Author notes

  1. P Panayiotidis & P Kotsi

    Present address: First Dept of Propedeutic Medicine, University of Athens, Laikon Hospital, Athens, Greece

  2. GL Palmisano

    Present address: Molecular Morphogenesis Lab, National Cancer Research Institute (IST) Genova, Largo R Benzi 10 16132, Genova, Italy

  3. C Rowntree and V Duke: These authors have contributed equally to this work

Authors and Affiliations

  1. Department of Academic Haematology Royal Free and University College of London, Pond Street, Royal Free Campus, London, UK

    C Rowntree, V Duke, AV Hoffbrand & L Foroni

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Rowntree, C., Duke, V., Panayiotidis, P. et al. Deletion analysis of chromosome 13q14.3 and characterisation of an alternative splice form of LEU1 in B cell chronic lymphocytic leukemia. Leukemia 16, 1267–1275 (2002). https://doi.org/10.1038/sj.leu.2402551

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