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Molecular and Cellular Biology, December 1999, p. 8536-8546, Vol. 19, No. 12
Institute of Microbiology and Immunology,
School of Life Science, National Yang-Ming University, Taipei,
Taiwan 11221, Republic of China
Received 16 June 1999/Returned for modification 17 July
1999/Accepted 17 August 1999
Nopp140 is thought to shuttle between nucleolus and cytoplasm.
However, the predominant nucleolar localization of Nopp140 homologues
from different species suggests that Nopp140 is also involved in events
occurring within the nucleolus. In this study, we demonstrated that the
largest subunit of RNA polymerase I, RPA194, was coimmunoprecipitated
with the human Nopp140 (hNopp140). Such an interaction is mediated
through amino acids 204 to 382 of hNopp140. By double
immunofluorescence, hNopp140 was colocalized with RNA polymerase I at
the rDNA (rRNA genes) transcription active foci in the nucleolus. These
results suggest that Nopp140 can interact with RNA polymerase I in
vivo. Transfected cells expressing the amino-terminal half of hNopp140,
hNopp140N382 (amino acids 1 to 382), displayed altered nucleoli with
crescent-shaped structures. This phenotype is reminiscent of the
segregated nucleoli induced by actinomycin D treatment, which is known
to inhibit rRNA synthesis. Consistently, the hNopp140N382 protein
mislocalized the endogenous RNA polymerase I and shut off cellular rRNA
gene transcription as revealed by an in situ run-on assay. These
dominant negative effects of the mutant hNopp140N382 suggest that
Nopp140 plays an essential role in rDNA transcription. Interestingly,
ectopic expression of hNopp140 to a very high level caused the
formation of a transcriptionally inactive spherical structure occupying the entire nucleolar area which trapped the RNA polymerase I, fibrillarin, and hNopp140 but excluded the nucleolin. The
mislocalizations of these nucleolar proteins after hNopp140
overexpression imply that Nopp140 may also play roles in maintenance of
nucleolar integrity.
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Human Nopp140, Which Interacts with RNA Polymerase
I: Implications for rRNA Gene Transcription and Nucleolar
Structural Organization

*
Corresponding author. Mailing address: Institute of
Microbiology and Immunology, School of Life Science, National Yang-Ming University, Taipei 11221, Taiwan. Phone: 886-2-2826-7113. Fax: 886-2-2821-2880. E-mail: yphcsl{at}ym.edu.tw.
Present address: Department of Biology, The Johns Hopkins
University, Baltimore, MD 21218.
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