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Functional cloning of SPIN-2, a nuclear anti-apoptotic protein with roles in cell cycle progression

Leukemia volume 16, pages 1507–1518 (2002)Cite this article

Abstract

The balance between hematopoietic cell viability and apoptosis is regulated by exogenous growth factors, however, the molecular mechanisms by which these trophic factors exert their effects remain obscure. A functional retroviral cDNA library-based screen was employed to identify genes that prevent growth factor withdrawal-mediated apoptosis in the myeloid progenitor cell 32Dcl3. This approach identified three classes of genes: those with known roles in apoptosis (bcl-XL and ornithine decarboxylase); genes previously identified but not linked directly to apoptotic signaling (O-linked N-acetylglucosamine transferase); and a previously uncharacterized gene we termed SPIN-2. In 32Dcl3 cells, expression of exogenous SPIN-2 provides 25% protection from apoptosis following growth factor withdrawal compared to controls which show ∼1–2% survival. SPIN-2 overexpression slows cell growth rates and increases the percentage of cells in G2/M (32% vs control cells at 12%). Immunolocalization studies indicate that myc-epitope tagged SPIN-2 proteins, which retain their anti-apoptotic function, reside in the nucleus, whereas a C-terminal deletion mutant that loses its anti-apoptotic activity is located in the cytoplasm. These studies suggest that SPIN-2 is a novel nuclear protein that functions to regulate cell cycle progression and this activity is related to the inhibition of apoptosis following the removal of essential growth factors.

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Acknowledgements

We thank Alan Friedman for the 32Dcl3 cells, Vijay Antharam for technical assistance and Drs Stratford May and Steven Baker for critical reading of the manuscript. The nucleotide sequence for the SPIN-2 gene has been submitted to the GenBank database under the Accession Number AF356353. A portion of this work was supported by a grant to BSF from the Howard Hughes Medical Institute, Postdoctoral Research Fellowship for Physicians.

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  1. Department of Pharmacology and Therapeutics, University of Florida, College of Medicine, Gainesville, FL, USA

    BS Fletcher & C Dragstedt

  2. Department of Neuroscience, University of Florida, College of Medicine, Gainesville, FL, USA

    L Notterpek

  3. Departments of Molecular Pharmacology and Microbiology and Immunology, Stanford University School of Medicine, Stanford, CA, USA

    GP Nolan

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Fletcher, B., Dragstedt, C., Notterpek, L. et al. Functional cloning of SPIN-2, a nuclear anti-apoptotic protein with roles in cell cycle progression. Leukemia 16, 1507–1518 (2002). https://doi.org/10.1038/sj.leu.2402557

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