The T/t-complex has an important role in the genetic control of cellular commitment in normal early development. Several important new studies make the recessive -lethal mutations more accessible to molecular analysis: (1) two of them have been mapped within the well characterized MHC; 12 is between H-2D and TL, and w5 is genetically inseparable from H-2K. (2) 185 kb of the H-2K region of -haplotypes has been cloned in cosmids and restriction mapped. Preliminary results indicate this region contains several early embryo-expressed sequences. (3) the structural gene for a -associated cell surface antigen, gp87, has two copies physically mapped, one in the Qa region and one between the two H-2K genes. We intend to use this recent information to try to identify and structurally characterize interesting early embryo-expressed genes including w5 and 12. The overall approaches we propose are: 1) to search the H-2K region cosmids for embryo expressed genes using cDNA from embryonal carcinoma cells, and to use unique copy probes from the genomic cosmids to probe available cDNA libraries from normal early embryos. In an effort to specifically identify w5, we propose to compare its H-2K region structure to that of a revertent. 2) The second approach is to structurally study the organization of the D-TL region of -haplotypes where the 12 gene is located. We will use CHEF gel analysis and existing recombinants to refine the map position of 12. 3) Finally, we will continue the molecular analysis of the -associated glycoprotein antigen """"""""glycoproteins 87"""""""" by retrieving and sequencing clones from cDNA libraries. We will study the expression of interesting genes identified using a variety of techniques including in situ hybridization to cut sections of embryos and in the case of the mutants, transgenic embryo rescue. Our long term objective is to understand the molecular mechanisms and genetic control of cellular commitment in normal early development, and noncommitment in embryonal tumors.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD030658-21
Application #
2025466
Study Section
Mammalian Genetics Study Section (MGN)
Project Start
1987-01-01
Project End
1998-12-31
Budget Start
1997-01-01
Budget End
1998-12-31
Support Year
21
Fiscal Year
1997
Total Cost
Indirect Cost
Name
University of Texas Austin
Department
Zoology
Type
Schools of Arts and Sciences
DUNS #
City
Austin
State
TX
Country
United States
Zip Code
78712
Saccomanno, L; Loushin, C; Jan, E et al. (1999) The STAR protein QKI-6 is a translational repressor. Proc Natl Acad Sci U S A 96:12605-10
Wu, J; Zhou, L; Tonissen, K et al. (1999) The quaking I-5 protein (QKI-5) has a novel nuclear localization signal and shuttles between the nucleus and the cytoplasm. J Biol Chem 274:29202-10
Venables, J P; Vernet, C; Chew, S L et al. (1999) T-STAR/ETOILE: a novel relative of SAM68 that interacts with an RNA-binding protein implicated in spermatogenesis. Hum Mol Genet 8:959-69
Vernet, C; Abe, K; Artzt, K (1998) Genetic mapping of 10 microsatellites in the t complex region of mouse chromosome 17. Mamm Genome 9:472
Rennebeck, G; Lader, E; Fujimoto, A et al. (1998) Mouse Brachyury the Second (T2) is a gene next to classical T and a candidate gene for tct. Genetics 150:1125-31
Rennebeck, G; Kleymenova, E V; Anderson, R et al. (1998) Loss of function of the tuberous sclerosis 2 tumor suppressor gene results in embryonic lethality characterized by disrupted neuroepithelial growth and development. Proc Natl Acad Sci U S A 95:15629-34
Vernet, C; Artzt, K (1997) STAR, a gene family involved in signal transduction and activation of RNA. Trends Genet 13:479-84
Zorn, A M; Grow, M; Patterson, K D et al. (1997) Remarkable sequence conservation of transcripts encoding amphibian and mammalian homologues of quaking, a KH domain RNA-binding protein. Gene 188:199-206
Hardy, R J; Loushin, C L; Friedrich Jr, V L et al. (1996) Neural cell type-specific expression of QKI proteins is altered in quakingviable mutant mice. J Neurosci 16:7941-9
Ebersole, T A; Chen, Q; Justice, M J et al. (1996) The quaking gene product necessary in embryogenesis and myelination combines features of RNA binding and signal transduction proteins. Nat Genet 12:260-5

Showing the most recent 10 out of 16 publications