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Nucleic Acids Research, 1984, Vol. 12, No. 7 3201-3217
© 1984

MOLECULAR BIOLOGY

Physical structure of gene-sized chromatin from the protozoan Oxytricha*

A.P. Butler+, T.J. Laughlin§, C.L. Cadilla, J.M. Henry and D.E. Olins

University of Tennessee, Oak Ridge Graduate School of Biomedical Sciences and Biology Division, Oak Ridge National Laboratory Oak Ridge, TN 37830, USA

Received November 14, 1983. Revised March 5, 1984. Accepted March 5, 1984.

Oxytricha nova is a hypotrichous ciliate containing a transcriptionally active macronucleus and a transcriptionally inactive micronucleus. Two-dimensional gel electrophores is shows that macronuclei contain a normal complement of inner histones. However, despite extensive efforts, no classical H1-like protein has been detected. Micrococcal nuclease digestion indicates a nucleosome repeat length of approximately 220 bp for macronuclear chromatin. Thermal denaturation profiles of macronuclear chromatin in 0.2 mM EDTA display four transitions at about 46, 57, 64, and 79°C. The lowest of these shifts to higher temperature as the ionic strength is raised to 3-5 mM Na phosohate. These results are consistent with the absence of H1 and a nucleosome repeat of 220–230 bp. Circular dichroism (CD) results aqree with these findings. By contrast, micronuclear chromatin displays a much smaller premelt and a more suppressed DNA CD signal at 285 nm, consistent with a micronuclear chromatin repeat of 165–185 bp as determined by micrococcal nuclease digestion.

*Research sponsored by the Office of Health and Environmental Research, U.S. Department of Energy, under contract W-7405-eng-26 with Union Carbide Corporation

+Present address: The University of Texas System Cancer Center, Science Park–Research Division, P.O. Box 389, Smithville, TX 78957, USA

§Present address: Southern Research Institute, 2000 Ninth Avenue S., Birmingham, AL 35255, USA


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Home page
 Genes Dev.Home page
C M Price and T R Cech
Telomeric DNA-protein interactions of Oxytricha macronuclear DNA.
Genes & Dev., October 1, 1987; 1(8): 783 - 793.
[Abstract] [PDF]


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