Non-Mendelian, heritable blocks to DNA rearrangement are induced by loading the somatic nucleus of Tetrahymena thermophila with germ line-limited DNA.
Site-specific DNA deletion occurs at thousands of sites within the genome during macronuclear development of Tetrahymena thermophila. These deletion elements are usually not detected in macronuclear chromosomes. We have interfered with the normal deletion of two of these elements, the adjacent M and R elements, by loading vegetative macronuclei with these elements prior to sexual conjugation. Transformed cell lines containing the exogenous M or R element, carried on high-copy-number vectors containing genes encoding rRNA within parental (old) macronuclei, consistently failed to excise chromosomal copies of the M or R element during formation of new macronuclei. Little or no interference with the deletions of adjacent elements or of unlinked elements was observed. The micronucleus (germ line)-limited region of each element was sufficient to inhibit specific DNA deletion. This interference with DNA deletion usually is manifested as a cytoplasmic dominant trait: deletion elements present in the old macronucleus of one partner of a mating pair were sufficient to inhibit deletion occurring in the other partner. Remarkably, the failure to excise these elements became a non-Mendelian, inheritable trait in the next generation and did not require the high copy number of exogenously introduced elements. The introduction of exogenous deletion elements into parental macronuclei provides us with an epigenetic means to establish a heritable pattern of DNA rearrangement.
Chalker
D L
DL
Division of Basic Sciences, Fred Hutchinson Cancer Research Center, Seattle, Washington 98104, USA. dchalker@fred.fhcrc.org
Yao
M C
MC
D000818:Animals; D001483:Base Sequence; D002467:Cell Nucleus / Q000378:metabolism; D003433:Crosses, Genetic; D017931:DNA Primers; D016054:DNA, Protozoan / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D048631:Micronucleus, Germline / Q000502:physiology; D008957:Models, Genetic; D008969:Molecular Sequence Data; D010957:Plasmids; D016133:Polymerase Chain Reaction; D017384:Sequence Deletion; D016808:Tetrahymena thermophila / Q000235:genetics* / Q000378:metabolism* / Q000502:physiology; D014170:Transformation, Genetic
D017931:DNA Primers; D016054:DNA, Protozoan