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The mitotic protein p62 is localized to the nucleolus during interphase.

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Call No.	QP625.N87 W37 2000
Author	Warner, Anne K.
Title	The mitotic protein p62 is localized to the nucleolus during interphase.
Date	2000
Abstract	Mitotic apparatuses from sea urchin embryos contain a protein (p62), previously shown to be required for mitotic progression. This protein localizes to the mitotic apparatus during cell division in urchin embryos and mammalian tissue culture cells. In this thesis new information about p62 is presented. Immunofluorescence microscopy revealed that p62 is located in the nucleus of mammalian cells during interphase and is highly concentrated in nucleoli. A fusion protein composed of full-length p62 and green fluorescent protein was generated and this also localized to nucleoli when expressed in COS-7 cells in culture. To identify the domain important for nucleolar localization during interphase, specific domains of p62 alone, or in combination with each other or with β-galactosidase were fused to green fluorescent protein. Following confirmation of the fusion constructs by sequence analysis, the constructs were expressed in mammalian cells, expression was confirmed by immunoblotting, and the fusion proteins were localized via fluorescence microscopy. The data demonstrated that the C-terminal domain of p62 (amino acid residues 321-411) is both necessary and sufficient for the nuclear localization and nucleolar binding of p62 that was observed during interphase. The localization of the fusion protein composed of full-length p62 and green fluorescent protein throughout mitosis in mammalian cells was determined by fluorescence microscopy. The data unexpectedly demonstrated that during mitosis this fusion protein is excluded from the chromosomes and is evenly distributed throughout the cytoplasm until telophase. The fusion protein was localized to punctate fluorescent particles in the nucleoplasm during telophase, and these particles appeared to coalesce into the reforming nucleoli. In addition, immunoblots containing samples from various human tissues were probed with p62 antiserum. The p62 antiserum recognized a protein of approximately 118 kDa in human testicular tissue, a tissue type with a significant amount of cell division. The p62 antiserum did not recognize proteins in any of the other human tissue samples tested. lmmunohistochemistry of human testicular tissue sections with p62 antiserum revealed staining in the nuclei of spermatocytes.
Description	1 online resource (x, 208 leaves) : illustrations (some color)
Format	application/pdf
Advisor	Sloboda, Roger D.
Subject	Nucleoproteins. ; Mitosis. ; Nucleolus. ; Molecular biology.
College	Dartmouth College -- Department of Biological Sciences.
Issuing Body	Dartmouth Dissertations are made available through the Dartmouth Digital Library.
Degree	Thesis (Ph.D)--Dartmouth College.
Copyright	In Copyright. This material is protected by copyright and/or related rights. You are free to use this material in any way that is permitted by copyright law that applies to your use. For other uses, you need to obtain permission from the rights-holder(s). http://rightsstatements.org/page/InC/1.0/
DOI	https://doi.org10.1349/ddlp.2913

Title	warner2000 1
Full Text	The Mitotic Protein p62 is Localized to the Nucleolus During Interphase A Thesis Submitted to the Faculty in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Biology by Anne K. Warner DARTMOUTH COLLEGE Hanover, New Hampshire June, 2000 Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.

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The mitotic protein p62 is localized to the nucleolus during interphase.

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