A proposal is made to analyze, by genetic and molecular techniques, a human gene whose expression is essential for normal neuroectodermal development. The research undertaken in this proposal will be directed towards the analysis of a human developmental gene, incontinentia pigmenti (IP). IP is an X-linked dominant, hemizygote lethal disorder. IP carrier females are identifiable and present with characteristic neuroectodermal developmental abnormalities. The IP locus has been tentatively localized to Xp11.21. This proposal provides for analysis of the IP locus by linkage analysis using collected genomic DNA samples from multigenerational IP families and characterized human X chromosomal probes capable of detecting restriction fragment length polymorphisms (RFLP's). An assemble collection of somatic cell hybrids containing rearranged X chromosomes will be used to isolated and map cloned X chromosomal fragments capable of detecting RFLP's, to the IP region. We propose to molecularly delineate the IP locus by identifying breakpoints of rearranged X chromosomes associated with the IP phenotype. Constructed somatic cell hybrids containing IP X/autosomal translocations will be used to identify X chromosomal overlaps, deletions, or breakpoint heterogeneity among the IP translocations by Southern blot analysis and pulse field gradient electrophoretic restriction mapping techniques. X chromosomal DNA fragments from the delineated Ip locus will be cloned. Unique, highly conserved regions from the IP locus will provide DNA fragment candidates for the isolation of the IP gene. These analyses will provide information to further delineate the genetic and chromosomal localization of a human developmental gene and provide the necessary starting point for cloning the IP gene. A more thorough understanding of neuroectodermal development, human developmental processes, genes and gen products necessary for normal completion of developmental process, and mutations that disrupt these processes should result from these studies.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS030771-07
Application #
2268740
Study Section
Mammalian Genetics Study Section (MGN)
Project Start
1991-09-24
Project End
1996-07-31
Budget Start
1994-09-01
Budget End
1996-07-31
Support Year
7
Fiscal Year
1994
Total Cost
Indirect Cost
Name
University of Michigan Ann Arbor
Department
Pediatrics
Type
Schools of Medicine
DUNS #
791277940
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109
Pasteris, N G; Buckler, J; Cadle, A B et al. (1997) Genomic organization of the faciogenital dysplasia (FGD1; Aarskog syndrome) gene. Genomics 43:390-4
Gorski, J L; Bialecki, M D; McDonald, M T et al. (1996) Cosmids map two incontinentia pigmenti type 1 (IP1) translocation breakpoints to a 180-kb region within a 1.2-Mb YAC contig. Genomics 35:338-45
Olson, M F; Pasteris, N G; Gorski, J L et al. (1996) Faciogenital dysplasia protein (FGD1) and Vav, two related proteins required for normal embryonic development, are upstream regulators of Rho GTPases. Curr Biol 6:1628-33
Pasteris, N G; Gorski, J L (1995) An intragenic TaqI polymorphism in the faciogenital dysplasia (FGD1) locus, the gene responsible for Aarskog syndrome. Hum Genet 96:494
Pasteris, N G; de Gouyon, B; Cadle, A B et al. (1995) Cloning and regional localization of the mouse faciogenital dysplasia (Fgd1) gene. Mamm Genome 6:658-61
Miller, A P; Gustashaw, K; Wolff, D J et al. (1995) Three genes that escape X chromosome inactivation are clustered within a 6 Mb YAC contig and STS map in Xp11.21-p11.22. Hum Mol Genet 4:731-9
Pasteris, N G; Cadle, A; Logie, L J et al. (1994) Isolation and characterization of the faciogenital dysplasia (Aarskog-Scott syndrome) gene: a putative Rho/Rac guanine nucleotide exchange factor. Cell 79:669-78
Burright, E N; Gorski, J L (1993) Interspersed repetitive sequence (IRS)-PCR amplification of pulsed-field gel fractionated DNA to derive markers from the incontinentia pigmenti 1 (IP1) locus. Nucleic Acids Res 21:175-6
Gorski, J L; Burright, E N (1993) The molecular genetics of incontinentia pigmenti. Semin Dermatol 12:255-65
Pasteris, N G; Bialecki, M D; Gorski, J L (1993) YAC subclone contig assembly by serial interspersed repetitive sequence (IRS)-PCR product hybridizations. Nucleic Acids Res 21:5275-6

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