Abstract
Nuclear matrix is a complex intranuclear network supposed to be involved in the various nuclear functions. In order to identify the nuclear matrix proteins, we isolated a cDNA clone from a human placenta cDNA library. This clone was partially represented a known cDNA clone HA1237. HA1237 encoded a 631-amino-acid peptide, which we designated NXP-1. NXP-1 was related to yeast Rad21/Scc1/Mcd1, Xenopus XRAD21, and mouse PW29, and identical with HR21spA isolated from a human testis cDNA library. We developed a polyclonal antibody to the purified NXP-1 bacterially expressed as a fusion protein with GST. Western blot analysis with anti-NXP-1 polyclonal antibody showed nuclear matrix localization of NXP-1 in HeLa cells. Indirect immunofluorescence staining also showed nuclear and nuclear matrix localization of the NXP-1. Results of in vitro binding assays employing nuclear matrix preparations indicated that the N-terminal region (16-128 amino acid) of NXP-1 has an important role in nuclear matrix distribution.
Copyright 2000 Academic Press.
MeSH terms
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Amino Acid Sequence
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Animals
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Apoptosis Regulatory Proteins
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Cell Cycle Proteins / chemistry*
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Cell Nucleus / metabolism*
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Chromosomal Proteins, Non-Histone
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DNA-Binding Proteins / chemistry*
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DNA-Binding Proteins / genetics*
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DNA-Binding Proteins / metabolism
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Erythroid-Specific DNA-Binding Factors
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Female
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Gene Library
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HeLa Cells
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Humans
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Liver / metabolism
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Male
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Mice
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Mice, Inbred BALB C
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Nuclear Matrix / metabolism*
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Nuclear Proteins / chemistry*
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Nuclear Proteins / genetics*
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Nuclear Proteins / metabolism
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Phosphoproteins / chemistry*
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Placenta / metabolism
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Pregnancy
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Rats
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / metabolism
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Saccharomyces cerevisiae / metabolism
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Saccharomyces cerevisiae Proteins*
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Sequence Deletion
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Testis / metabolism
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Transcription Factors / chemistry*
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Transcription Factors / genetics*
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Transcription Factors / metabolism
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Xenopus
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Xenopus Proteins*
Substances
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Apoptosis Regulatory Proteins
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Cell Cycle Proteins
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Chromosomal Proteins, Non-Histone
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DNA-Binding Proteins
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Erythroid-Specific DNA-Binding Factors
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MCD1 protein, S cerevisiae
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Nuclear Proteins
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Phosphoproteins
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RAD21 protein, human
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Rad21 protein, Xenopus
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Rad21 protein, mouse
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Recombinant Fusion Proteins
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Saccharomyces cerevisiae Proteins
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Transcription Factors
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Xenopus Proteins