Abstract
EcoRI subclones, designated as 50E1 and 50E4, were independently obtained from a cosmid clone previously mapped to the centromeric region of human chromosome 8. Southern blot hybridization analyses suggested that both subclones contain repetitive DNA sequences different from the chromosome 8 specific alphoid DNA. DNA sequence analysis of the 704 bp insert of 50E1 and the 1, 962 bp insert of 50E4 revealed that both inserts contained tandemly repeated units of ∼220 bp. Fluorescence in situ hybridization studies confirmed these two subclones to be specifically located on the centromeric region of chromosome 8. A 220 bp consensus sequence, derived from nine monomeric repeats, showed no significant homology to alphoid consensus sequences or to other currently known human centromeric DNA sequence. Furthermore, no significant homology was found with any other DNA sequence deposited in the EMBL or GenBank databases, indicating that this chromosome 8 specific repetitive DNA sequence is novel. From slot blot experiments it was estimated that 0.013% of the human genome comprises 1,750 of these monomeric repeats, residing on the centromeric region of chromosome 8 in tandem array(s).
Similar content being viewed by others
References
Choo KH, Vissel B, Nagy A, Earle E, Kalitsis P (1991) A survey of the genomic distribution of alpha satellite DNA on all the human chromosomes, and derivation of a new consensus sequence. Nucleic Acids Res 19: 1179–1182
Corneo G, Ginelli E, Polli E (1967) A satellite DNA isolated from human tissue. J Mol Biol 23: 619–622
Corneo G, Ginelli E, Polli E (1970) Repeated sequences in human DNA. J Mol Biol 48: 319–327
Corneo G, Ginelli E, Polli E (1971) Renaturation properties and localization in heterochromatin of human satellite DNAs. Biochim Biophys Acta 247: 528–534
Donlon TA, Bruns GA, Latt SA, Mulholland J, Wyman AR (1987) A chromosome 8 enriched alphoid repeat. (Human gene mapping 9) Cytogenet Cell Genet 46: 607
Fan YS, Sasi R, Lee C, Court D, Lin CC (1992) Mapping of 50 cosmid clones isolated from a flow-sorted human X chromosome library by fluorescence in situ hybridization. Genomics 14: 542–545
Ge Y, Wagner MJ, Siciliano M, Wells DE (1992) Sequence, higher order repeat structure, and long-range organization of alpha satellite DNA specific to human chromosome 8. Genomics 13: 585–593
Gosden JR, Mitchell AR, Buckland RA, Clayton RP, Evans HJ (1975) The location of four human satellite DNAs on human chromosomes. Exp Cell Res 92: 148–158
Grady DL, Ratliff RL, Robinson DL, McCanlies EC, Meyne J, Moyzis RK (1992) Highly conserved repetitive DNA sequences are present at human centromeres. Proc Natl Acad Sci USA 89: 1695–1699
Greig GM, Willard HF, (1992) β satellite DNA: Characterization and localization of two subfamilies from the distal and proximal short arms of the human acrocentric chromosomes. Genomics 12: 573–580
Hadlaczky G, Praznovszky T, Cserpan I, Kereso J Peterfy M, Kelemen I, Atalay E, Szeles A, Szelei J Tubak V (1991) Centromere formation in mouse cells co-transformed with human DNA and a dominant marker gene. Proc Natl Acad Sci USA 88: 8106–8110
Hamkalo BA, Narayanswami S, Lundgren K (1989) Localization of nucleic acid sequences by EM in situ hybridization using colloidal gold labels. Am J Anat 185: 197–204
Higgins MJ, Wang H, Shtromas I, Haliotis T, Roder JC, Holden JJA, White BN (1985) Organization of a repetitive human 1.8 kb KpnI sequence localized in the heterochromatin of chromosome 15. Chromosoma 93: 77–86
Jabs EW, Persico MG (1987) Characterization of human centromeric regions of specific chromosomes by means of alphoid DNA sequences. Am J Hum Genet 41: 374–390
Jabs EW, Goble CA, Cutting CR (1989) Macromolecular organization of human centromeric regions reveals high-frequency, polymorphic macro DNA repeats. Proc Natl Acad Sci USA 86: 202–206
Lin CC, Meyne J, Sasi R, Bowen P, Unger T, Tainaka T, Hadro TA, Hoo JJ (1990) Determining the origins and structural aberrations of small marker chromosomes in two cases of 45, X/46, X, +mar by use of chromosome-specific DNA probes. Am J Med Genet 37: 71–78
Lin CC, Sasi R, Fan YS, Chen ZQ (1991) New evidence for tandem chromosome fusions in the karyotypic evolution of Asian muntjacs. Chromosoma 101: 19–24
Mandel P, Metais P, Cuny S (1950) Les quantites de DNA per leucocyte chez diverses especes de mammiferes. CR Acad Sci [III] 231: 1172–1174
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY
Manuelidis L, Wu JC (1978) Homology beteween human and simian repeated DNA. Nature 276: 92–94
McCutchan T, Hsu H, Thayer RE, Singer MF (1982) Organization of African green monkey DNA at junction between α-satellite and other DNA sequences. J Mol Biol 157: 195–211
Mitchell AR, Gosden JR, Miller DA (1985) p82H: a cloned sequence of the alphoid repeated DNA family found at the centromeres of all human chromosomes. Chromosoma 92: 369–377
Muro Y, Masumoto H, Yoda K, Nozaki N, Ohashi M, Okazaki T (1992) Centromere protein B assembles human centromeric α-satellite DNA at the 17 bp sequence. CENP-B box. J Cell Biol 116: 585–596
Prosser J, Frommer M, Paul C, Vincent PC, (1986) Sequence relationships of three human satellite DNAs. J Mol Biol 187: 145–155
Vissel B, Choo KH (1987) Human alpha satellite DNA—consensus sequence and conserved regions. Nucleic Acids Res 15: 6751–6752
Vissel B, Nagy A, Choo KHA (1992) A satellite III sequence shared by human chromosomes 13, 14, and 21 that is contiguous with α satellite DNA. Cytogenet Cell Genet 61: 81–86
Warburton PE, Willard HF (1990) Genomic analysis of sequence variation in tandemly repeated DNA, evidence for localized homogeneous sequence domains within arrays of α-satellite DNA. J Mol Biol 216: 3–16
Waye JS, Willard HF (1987) Nucleotide sequence heterogeneity of alpha satellite repetitive DNA: a survey of alphoid sequences from different human chromosomes. Nucleic Acids Res 15: 7549–7569
Waye JS, Willard HF (1989) Human β-satellite DNA: Genomic organization and sequence definition of a class of highly repetitive tandem DNA. Proc Natl Acad Sci USA 86: 6250–6254
Weier H-UG, Kleine H-D, Gray JW (1991) Labelling of the centromeric region on human chromosome 8 by in situ hybridization. Hum Genet 87: 489–494
Willard HF, Waye JS (1987) Chromosome-specific subsets of human alpha satellite DNA: Analysis of sequence divergence within and between chromosomal subsets and evidence for an acestral pentameric repeat. J Mol Evol 25: 207–214
Wu JC, Manuelidis L (1980) Sequence definition and organization of a human repeated DNA. J Mol Biol 142: 363–386
Author information
Authors and Affiliations
Additional information
Communicated by: J.B. Rattner
Rights and permissions
About this article
Cite this article
Lin, C.C., Sasi, R., Fan, Y.S. et al. Isolation and identification of a novel tandemly repeated DNA sequence in the centromeric region of human chromosome 8. Chromosoma 102, 333–339 (1993). https://doi.org/10.1007/BF00661276
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00661276