Abstract

The goal of the research is to characterize the structure and function of the human centromeric region. The centromere is the cytologically visible and genetically defined region of human chromosomes which is responsible for proper segregation of chromosomes in both mitosis and meiosis. These processes involve specialized chromosomal DNA sequences which interact specifically with cellular components. To investigate those DNA sequences that are required for centromeric activity the following research plan is proposed: 1. Sequences from the regions around the centromere of specific human chromosomes will be isolated by screening chromosome sorted libraries with 308, a human DNA sequence from chromosome 6, but homologous to centromeres of all human chromosomes. DNA from centromeric regions flanking the 308 related sequences will be obtained from screening recombinants for overlapping sequences. In this way unique as well as repetitive sequences will be obtained. 2. Centromeric sequences will be characterized to determine chromosomal localization by in situ hybridization to metaphase chromosomes and organization by Southern blot analysis and nucleotide sequence analysis. 3. To test for centromeric function, human DNA sequences will be inserted into viral vectors which usually integrate into chromosomes and transform mammalian cells. In these cells, sequences which function as centromeres will permit vectors to be stably maintained and segregate correctly with each cell division. The extrachromosomal localization will be determined by in situ hybridization and the organization of these sequences will be analyzed by Southern blot analysis. 4. Robertsonian translocation chromosomes will be analyzed with respect to the organization of their centromeric regions by using in situ hybridization, Southern blot analysis, and nucleotide sequence analysis. 5. Centromeric DNA probes will be used as cytological markers in in situ hybridization studies to determine the arrangement of chromosomes in the interphase nucleus.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD022819-03
Application #
3322707
Study Section
Mammalian Genetics Study Section (MGN)
Project Start
1986-08-01
Project End
1991-07-31
Budget Start
1988-08-01
Budget End
1989-07-31
Support Year
3
Fiscal Year
1988
Total Cost
Indirect Cost

  Institution

Name
Johns Hopkins University
Department
Type
Schools of Medicine
DUNS #
045911138
City
Baltimore
State
MD
Country
United States
Zip Code
21218

  Publications

Patton, S; Gendler, S J; Spicer, A P (1995) The epithelial mucin, MUC1, of milk, mammary gland and other tissues. Biochim Biophys Acta 1241:407-23
Lee, C; Li, X; Jabs, E W et al. (1995) Human gamma X satellite DNA: an X chromosome specific centromeric DNA sequence. Chromosoma 104:103-12
Bowcock, A M; Gerken, S C; Barnes, R I et al. (1993) The CEPH consortium linkage map of human chromosome 13. Genomics 16:486-96
Jabs, E W; Tuck-Muller, C M; Anhalt, G J et al. (1993) Cytogenetic survey in systemic sclerosis: correlation of aneuploidy with the presence of anticentromere antibodies. Cytogenet Cell Genet 63:169-75
Cooper, L F; Coss, C A; Jabs, E W (1992) Reevaluation of the origin of a marker chromosome in a patient with 47,XX,r(13)(p11q34), + mar by molecular cytogenetics. Clin Genet 42:323-5
Holden, K R; Jabs, E W; Sponseller, P D (1992) Roberts/pseudothalidomide syndrome and normal intelligence: approaches to diagnosis and management. Dev Med Child Neurol 34:534-9
Stetten, G; Blakemore, K J; Courter, A M et al. (1992) Prenatal identification of small mosaic markers of different chromosomal origins. Prenat Diagn 12:83-91
Jabs, E W; Tuck-Muller, C M; Cusano, R et al. (1991) Studies of mitotic and centromeric abnormalities in Roberts syndrome: implications for a defect in the mitotic mechanism. Chromosoma 100:251-61
Jabs, E W; Warren, A C; Taylor, E W et al. (1991) Alphoid DNA polymorphisms for chromosome 21 can be distinguished from those of chromosome 13 using probes homologous to both. Genomics 9:141-6
Blanche, H; Zoghbi, H Y; Jabs, E W et al. (1991) A centromere-based genetic map of the short arm of human chromosome 6. Genomics 9:420-8

Showing the most recent 10 out of 21 publications