Interspersion of mouse satellite deoxyribonucleic acid sequences.

P J Stambrook
Author Information

Abstract

DNA sequences with homology to the major (A + T)-rich mouse satellite component were localized in CsCl gradients by hybridization with a labeled satellite cRNA probe. Although, as expected, most of the hybridization was to DNA in the satellite-rich shoulder, substantial radioactive cRNA hybridized with DNA from denser regions of the gradient. Further examination revealed that hybridization to main-band DNA was not due to physical trapping of satellite DNA in the gradient, and melting experiments argue that the associated radioactivity was due to true RNA/DNA hybridization. Nearest-neighbor analysis of hybridized [alpha-32P]CTP-labeled l-strand cRNA indicates that hybridization to main-band DNA is by the satellite cRNA and not a contaminant. Together, these data argue that mouse satellite-like sequences are interspersed within the main-band fraction of DNA. For the support of this contention, total mouse DNA, purified main-band DNA, and purified satellite DNA were digested with EcoRI, sedimented in a sucrose gradient, and hybridized with labeled satellite cRNA. Mouse satellite DNA is not cleaved with EcoRI, so that purified EcoRI-digested satellite DNA sediments as a high molecular weight component. When total mouse DNA is digested with EcoRI, the majority of satellite-like sequences remain as high molecular weight DNA; however, significant amounts of satellite-like sequences sediment with the bulk of the lower molecular weight digested DNA, lending further credence to the argument that satellite-like sequences are interspersed with main-band DNA.

Grants

  1. GM 22248/NIGMS NIH HHS
  2. GM 29463/NIGMS NIH HHS

MeSH Term

Animals
Base Sequence
Centrifugation, Density Gradient
DNA
DNA Restriction Enzymes
DNA, Satellite
Kinetics
Liver
Mice
Mice, Inbred C57BL
Mice, Inbred CBA
Nucleic Acid Hybridization

Chemicals

DNA, Satellite
DNA
DNA Restriction Enzymes

Word Cloud

Created with Highcharts 10.0.0DNAsatellitesequencesmousehybridizationcRNAmain-bandsatellite-likehybridizedgradientpurifieddigestedEcoRImolecularweightcomponentlabeleddueargueinterspersedtotalhighhomologymajor+T-richlocalizedCsClgradientsprobeAlthoughexpectedsatellite-richshouldersubstantialradioactivedenserregionsexaminationrevealedphysicaltrappingmeltingexperimentsassociatedradioactivitytrueRNA/DNANearest-neighboranalysis[alpha-32P]CTP-labeledl-strandindicatescontaminantTogetherdatawithinfractionsupportcontentionsedimentedsucroseMousecleavedEcoRI-digestedsedimentsmajorityremainhoweversignificantamountssedimentbulklowerlendingcredenceargumentInterspersiondeoxyribonucleicacid

Similar Articles

Cited By