We have isolated, cloned, and sequenced, from the genome of human (HeLa) cells, a 1.797 Kb EcoR1 satellite II DNA fragment that displayed partial identity to the Drosophila melanogaster transposable P-element. / The sequence analysis of this clone (pKS36) indicated that it has originated from the tandem amplification of pentameric repeats that were derived from satellite II and III canonical consensus sequence 5$ sp prime$ TTCCA 3$ sp prime$. A two-stage decay mechanism, based on the methylation and subsequent deamination of cytosine residues within the pentamers, was proposed to explain the non-random base substitutions that were observed in pKS36, as well as in other sequenced members of the satellite II and III DNA families. In addition, this two-stage decay model correlated with the Tag1 and Hinf1 polymorphisms that were observed between related satellite DNA members. / Clone pKS36 also contains a region of 49 bp devoid of satellite sequences that was found, by southern hybridization, to be present in pKS36 closely related (pKS36-like) but absent from more divergent (pKS36-related) cloned satellite II and III DNAs. pKS36-related satellite DNAs represent up to 2% of the genomes of HeLa and MeWo cells, and are organized mostly in tandem arrays of 1.8 Kb EcoR1, Kpn1, and Sau3A DNA fragments. pKS36-like satellite DNAs represent less than 1% of the genome of HeLa cells, and are found mainly organized as 1.65 Kb, 1.95 Kb and 3.6 Kb EcoR1 elements, though their Kpn1 and Sau3A distributions resemble that of pKS36-related satellite DNAs. Cell specific organization of satellite DNAs, that may be the result of chromosomal translocations inherent to cultured cells, was observed in the two human (HeLa and MeWo) cell lines. / The analysis, by southern hybridization, of satellite DNAs using field inversion gel electrophoresis revealed the presence, in HeLa cells, of satellite DNA clusters ranging from 150 Kb to 500 Kb in length. / Using rodent-human hybrid cell DNAs, the members of the pKS36 satellite II DNA family were found to reside mainly on human chromosomes 7, 12, 14, 15, 16, and 22.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.70317 |
| Date | January 1991 |
| Creators | Sol, Katia |
| Publisher | McGill University |
| Source Sets | Library and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada |
| Language | English |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Format | application/pdf |
| Coverage | Doctor of Philosophy (Department of Microbiology and Immunology.) |
| Rights | All items in eScholarship@McGill are protected by copyright with all rights reserved unless otherwise indicated. |
| Relation | alephsysno: 001277497, proquestno: AAINN74782, Theses scanned by UMI/ProQuest. |
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