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Vol. 10, Issue 12, 4091-4106, December 1999
Department of Biochemistry, University of Mississippi Medical
Center, Jackson, Mississippi 39216
Quiescent nuclei from differentiated somatic cells can reacquire
pluripotence, the capacity to replicate, and reinitiate a program of
differentiation after transplantation into amphibian eggs. The
replication of quiescent nuclei is recapitulated in extracts derived
from activated Xenopus eggs; therefore, we have exploited this cell-free system to explore the mechanisms that regulate
initiation of replication in nuclei from terminally differentiated Xenopus erythrocytes. We find that these nuclei lack
many, if not all, pre-replication complex (pre-RC) proteins. Pre-RC
proteins from the extract form a stable association with the chromatin of permeable nuclei, which replicate in this system, but not with the
chromatin of intact nuclei, which do not replicate, even though these
proteins cross an intact nuclear envelope. During extract incubation,
the linker histones H1 and H10 are removed from erythrocyte
chromatin by nucleoplasmin. We show that H1 removal facilitates the
replication of permeable nuclei by increasing the frequency of
initiation most likely by promoting the assembly of pre-RCs on
chromatin. These data indicate that initiation in erythrocyte nuclei
requires the acquisition of pre-RC proteins from egg extract and that
pre-RC assembly requires the loss of nuclear envelope integrity and is
facilitated by the removal of linker histone H1 from chromatin.
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 S. A. Grigoryev, T. Nikitina, J. R. Pehrson, P. B. Singh, and C. L. Woodcock Dynamic relocation of epigenetic chromatin markers reveals an active role of constitutive heterochromatin in the transition from proliferation to quiescence J. Cell Sci., December 1, 2004; 117(25): 6153 - 6162. [Abstract] [Full Text] [PDF]
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 S. Novak, F. Paradis, C. Savard, K. Tremblay, and M.-A. Sirard Identification of Porcine Oocyte Proteins That Are Associated with Somatic Cell Nuclei after Co-Incubation Biol Reprod, October 1, 2004; 71(4): 1279 - 1289. [Abstract] [Full Text] [PDF]
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S. A. Grigoryev, T. Nikitina, J. R. Pehrson, P. B. Singh, and C. L. Woodcock Dynamic relocation of epigenetic chromatin markers reveals an active role of constitutive heterochromatin in the transition from proliferation to quiescence J. Cell Sci., December 1, 2004; 117(25): 6153 - 6162. [Abstract] [Full Text] [PDF]
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