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

Leukemia. 2002 Aug;16(8):1507-18. doi: 10.1038/sj.leu.2402557.

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-X(L) 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 approximately 1-2% survival. SPIN-2 overexpression slows cell growth rates and increases the percentage of cells in G(2)/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.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3 Cells
  • Amino Acid Sequence
  • Animals
  • Apoptosis / genetics*
  • Cell Cycle / genetics*
  • Cell Cycle Proteins / genetics
  • Cell Cycle Proteins / isolation & purification*
  • Cell Cycle Proteins / physiology
  • Cell Line / cytology
  • Cell Line / metabolism
  • Cloning, Molecular
  • DNA, Complementary / genetics
  • Gene Expression Profiling
  • Gene Library
  • Genetic Vectors / genetics
  • HL-60 Cells / chemistry
  • Humans
  • Interleukin-3 / pharmacology
  • Jurkat Cells / cytology
  • Mice
  • Molecular Sequence Data
  • Myeloid Cells / cytology
  • Myeloid Cells / metabolism
  • Nuclear Proteins / genetics
  • Nuclear Proteins / isolation & purification*
  • Nuclear Proteins / physiology
  • RNA, Messenger / genetics
  • RNA, Neoplasm / genetics
  • Recombinant Fusion Proteins / physiology
  • Retroviridae / genetics
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • Structure-Activity Relationship
  • X Chromosome / genetics

Substances

  • Cell Cycle Proteins
  • DNA, Complementary
  • Interleukin-3
  • Nuclear Proteins
  • RNA, Messenger
  • RNA, Neoplasm
  • Recombinant Fusion Proteins
  • SPIN2A protein, human

Associated data

  • GENBANK/AF356353