Fifty years ago Haldene proposed the hypothesis that new mutation account for about one-third of cases of hemophilia and other X-linked """"""""genetic lethal"""""""" conditions. He also stated that in a stationary population mutations to hemophilia would have a life of two to four generations. We now have the opportunity to determine directly the accuracy of Haldane's theory. In this proposal we will screen by DNA analysis about 500 hemophilia A families, and we expect to characterize the mutant allele of the factor VIII:C gene in about 100 of these families. (Parenthetically, Haldane postulated the existence of just two hemophilia alleles in 1935). We believe this is possible because we have found the mutation in 16 of 80 mutant genes screened to date. Among the 100 alleles we will discover that 50 of these mutations occurrred de novo within two generations of the proband. We will then determine 1) the distribution of de novo mutations in these tow generations, 2) the types of mutations (point mutations vs. deletions) and their sex distribution, and 3) whether advanced paternal (or maternal) age is associated with various types of new mutations. In characterizing the molecular defect in many of these mutant factor VIII:C genes, we expect to gain significant new insights into structure-function relationship in the factor VIII:C protein. In addition, we will quantify the extent to which CpG dinucleotides are mutation sensitive and discover whether particular CpG residues are more sensitive to mutation than others. Preliminary data indicate that this should be a highly useful approach because CG-TG and CG-CA mutations have now been observed by us and Gitschier et al in seven mutant alleles at 5 CpG sites studied in exon of the factor VIII:C gene. Two instances of recurrent CGA-TGA nonsenses mutations have been found which account for four alleles among the 80 mutant genes studied by us. In summary, the work proposed should provide definitive answers to a number of improtant questions involving the generation of mutations in man.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL038165-03
Application #
3354221
Study Section
Mammalian Genetics Study Section (MGN)
Project Start
1987-04-01
Project End
1991-03-31
Budget Start
1989-04-01
Budget End
1990-03-31
Support Year
3
Fiscal Year
1989
Total Cost
Indirect Cost
Name
Johns Hopkins University
Department
Type
Schools of Medicine
DUNS #
045911138
City
Baltimore
State
MD
Country
United States
Zip Code
21218
Leaf, Alexander; Xiao, Yong Fu; Kang, Jing X et al. (2003) Prevention of sudden cardiac death by n-3 polyunsaturated fatty acids. Pharmacol Ther 98:355-77
Fakharzadeh, S S; Zhang, Y; Sarkar, R et al. (2000) Correction of the coagulation defect in hemophilia A mice through factor VIII expression in skin. Blood 95:2799-805
Neerman-Arbez, M; Antonarakis, S E; Honsberger, A et al. (1999) The 11 kb FGA deletion responsible for congenital afibrinogenaemia is mediated by a short direct repeat in the fibrinogen gene cluster. Eur J Hum Genet 7:897-902
Neerman-Arbez, M; Johnson, K M; Morris, M A et al. (1999) Molecular analysis of the ERGIC-53 gene in 35 families with combined factor V-factor VIII deficiency. Blood 93:2253-60
Antonarakis, S E (1998) Molecular genetics of coagulation factor VIII gene and haemophilia A. Haemophilia 4 Suppl 2:1-11
Neerman-Arbez, M; Antonarakis, S E; Blouin, J L et al. (1997) The locus for combined factor V-factor VIII deficiency (F5F8D) maps to 18q21, between D18S849 and D18S1103. Am J Hum Genet 61:143-50
Young, M; Inaba, H; Hoyer, L W et al. (1997) Partial correction of a severe molecular defect in hemophilia A, because of errors during expression of the factor VIII gene. Am J Hum Genet 60:565-73
Bi, L; Sarkar, R; Naas, T et al. (1996) Further characterization of factor VIII-deficient mice created by gene targeting: RNA and protein studies. Blood 88:3446-50
Antonarakis, S E; Kazazian, H H; Gitschier, J et al. (1995) Molecular etiology of factor VIII deficiency in hemophilia A. Adv Exp Med Biol 386:19-34
Antonarakis, S E (1995) Molecular genetics of coagulation factor VIII gene and hemophilia A. Thromb Haemost 74:322-8

Showing the most recent 10 out of 19 publications