Genetic analysis of the APAFI gene in male germ cell tumors Journal Article
| Authors: | Bala, S.; Oliver, H.; Renault, B.; Montgomery, K.; Dutta, S.; Rao, P.; Houldsworth, J.; Kucherlapati, R.; Wang, X.; Chaganti, R. S. K.; Murty, V. V. V. S. |
| Article Title: | Genetic analysis of the APAFI gene in male germ cell tumors |
| Abstract: | Cytogenetic and molecular analyses have shown that the chromosome band 12q22 is recurrently deleted in male germ cell tumors (GCTs), indicating the presence of a candidate tumor suppressor gene (TSG) in this region. To identify the TSG, we mapped the APAFI gene, a proapoptotic mammalian homologue of ced-4, to chromosomal band 12q22, that suggested that this might be the candidate deleted gene in GCTs. We further localized the gene between the polymorphic markers D12S1671 and D12S1082 at 12q22 to determine the role of APAFI in the pathogenesis of GCT, and we characterized its normal genomic structure and analyzed its alterations in GCTs. The APAFI gene comprises 27 exons, with the coding region spanning 26. The region containing APAFI was found to be deleted in GCT by fluorescence in situ hybridization analysis, but without evidence of coding sequence alterations. RT-PCR and Western blot analysis showed APAFI gene expression at detectable levels in all GCT cell lines analyzed. An aberrant-sized APAFI protein was seen in one cell line. This and 2 other cell lines carrying APAFI deletions also exhibited defects in dATP-mediated caspase-3 activation. Caspase-3 activity was effectively restored by addition of recombinant caspase-9 and APAFI proteins, and to a lesser extent by caspase-9 alone, but not by APAFI alone. These data do not support a TSG role for APAFI, but defects in other components of the apoptotic pathway that may be related to 12q22 deletion cannot be ruled out. (C) 2000 Wiley-Liss, Inc. |
| Keywords: | human tissue; unclassified drug; human cell; exon; genetics; exons; molecular genetics; proteins; metabolism; mammalia; apoptosis; protein; intron; introns; gene product; caspase 3; enzymology; pathology; caspase; caspases; gene mapping; tumor suppressor gene; biosynthesis; dna; molecular sequence data; messenger rna; protein synthesis; rna, messenger; nucleotide sequence; dna, neoplasm; testis tumor; testicular neoplasms; protein biosynthesis; base sequence; caspase 9; dna mutational analysis; chromosome deletion; germ cell tumor; polymorphism, genetic; hydrolysis; genetic marker; chromosome 12q; genetic markers; genetic polymorphism; chromosome map; germinoma; apoptotic protease activating factor 1; apoptotic protease-activating factor 1; chromosome banding pattern; humans; human; male; priority journal; article; casp3 protein, human; apaf1 protein, human; physical chromosome mapping |
| Journal Title: | Genes Chromosomes and Cancer |
| Volume: | 28 |
| Issue: | 3 |
| ISSN: | 1045-2257 |
| Publisher: | Wiley Periodicals, Inc |
| Date Published: | 2000-07-01 |
| Start Page: | 258 |
| End Page: | 268 |
| Language: | English |
| DOI: | 10.1002/1098-2264(200007)28:3<258::aid-gcc3>3.0.co;2-r |
| PUBMED: | 10862031 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Export Date: 18 November 2015 -- Source: Scopus |
Altmetric
Citation Impact
BMJ Impact Analytics
MSK Authors
Related MSK Work