The long term objective of this proposal is to produce and accurate physical map and complete molecular clones of human chromosome segment 18q21.3 and to use this to characterize human diseases mapping to 18q21.3. Six probes known to map 18q21.3 provide the initial toehold in this segment. A long-range physical map of this segment using contour- clamped homogeneous electric field electrophoresis (CHEF) and rare cutting restriction endonucleases will order these loci. However, long-range restriction maps have technical limitations and emphasize the need to obtain large contiguous molecular clones. This approximately 2 megabase segment constitutes a reasonably sized test case to obtain overlapping clones from a yeast artificial chromosome (YAC) library of the human genome. YAC isolates obtained with the six 18q21.3 probes will be assessed for their authenticity and further mapped. Distal ends of YAC genomic inserts will be rescued and used to rescreen the library to link clones of 18q21.3. Molecular clones of this region provide the framework to localize new genes reveal associations between geography and the regulation of neural and immune related genes. Follicular lymphomas translocate a new proto-oncogene, Bc1-2 from 18q21.3, into the immunoglobulin locus (14q32) resulting in a marked overproduction of Bc1-2 mRNA. However, CHEF analysis reveals that the Bc1-2 gene possesses and enormous approximately 350 Kb Intron II and that the deregulatory effects of translocation are transmitted across this distance. YAC technology provides the capability to clone this intron as a single fragment. Other genes located within this intron would be identified, cloned, and their effects upon neoplasia or other cell types assessed. The 18q- deletion syndrome is a congenital developmental abnormality manifesting as craniofacial abnormalities, mental retardation, and humoral immunodeficiency. The deleted segment common to these patients is band 18q21.3 18q- syndrome patients with interstitial deletions or constitutional translocations at 18q21.3 will be used to identify distinct breakpoints. This should determine whether a single gene is always altered or whether multigenic loss is responsible. The physical map and YAC clones of 18q21.3 open new avenues for assessing normal and pathologic gene regulation and serve as a model for approaching human genetic disease.
| Edwards, Amanda L; Wachter, Franziska; Lammert, Margaret et al. (2015) Cellular Uptake and Ultrastructural Localization Underlie the Pro-apoptotic Activity of a Hydrocarbon-stapled BIM BH3 Peptide. ACS Chem Biol 10:2149-57 |
| Barclay, Lauren A; Wales, Thomas E; Garner, Thomas P et al. (2015) Inhibition of Pro-apoptotic BAX by a noncanonical interaction mechanism. Mol Cell 57:873-886 |
| Zhang, Tejia; Walensky, Loren D; Saghatelian, Alan (2015) A nonapoptotic role for BAX and BAK in eicosanoid metabolism. ACS Chem Biol 10:1398-403 |
| Carpio, Marcos A; Michaud, Michael; Zhou, Wenping et al. (2015) BCL-2 family member BOK promotes apoptosis in response to endoplasmic reticulum stress. Proc Natl Acad Sci U S A 112:7201-6 |
| Zhang, Tejia; Barclay, Lauren; Walensky, Loren D et al. (2015) Regulation of mitochondrial ceramide distribution by members of the BCL-2 family. J Lipid Res 56:1501-10 |
| Lee, Susan; Braun, Craig R; Bird, Gregory H et al. (2014) Photoreactive stapled peptides to identify and characterize BCL-2 family interaction sites by mass spectrometry. Methods Enzymol 544:25-48 |
| Walensky, Loren D; Bird, Gregory H (2014) Hydrocarbon-stapled peptides: principles, practice, and progress. J Med Chem 57:6275-88 |
| Bird, Greg H; Gavathiotis, Evripidis; LaBelle, James L et al. (2014) Distinct BimBH3 (BimSAHB) stapled peptides for structural and cellular studies. ACS Chem Biol 9:831-7 |
| Edwards, Amanda L; Gavathiotis, Evripidis; LaBelle, James L et al. (2013) Multimodal interaction with BCL-2 family proteins underlies the proapoptotic activity of PUMA BH3. Chem Biol 20:888-902 |
| Walensky, Loren D (2013) Protein-protein interactions: A PUMA mechanism unfolds. Nat Chem Biol 9:141-3 |
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