Abstract
NAP-1, a protein first isolated from mammalian cells, can introduce supercoils into relaxed circular DNA in the presence of purified core histones. Based on its in vitro activity, it has been suggested that NAP-1 may be involved in nucleosome assembly in vivo. We isolated a cDNA clone encoding a soybean NAP-1 homolog, SNAP-1. The SNAP-1 cDNA contains an open reading frame of 358 amino acid residues with a calculated molecular weight of 41 kDa. The deduced amino acid sequence of SNAP-1 shares sequence similarity with yeast NAP-1 (38%) and human hNRP (32%). Notable features of the deduced sequence are two extended acidic regions thought to be involved in histone binding. SNAP-1 expressed in Escherichia coli induces supercoiling in relaxed circular DNA, suggesting that SNAP-1 may have nucleosome assembly activity. The specific activity of SNAP-1 is comparable to that of HeLa NAP-1 in an in vitro assay. Western analysis reveals that SNAP-1 is expressed in the immature and young tissues that were examined, while mature tissues such as old leaves and roots, show very little or no expression. NAP-1 homologs also appear to be present in other plant species.
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References
Adams CC, Workman JL (1993) Nucleosome displacement in transcription. Cell 72:305–308
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 72:248–254
Chen H, Li B, Workman JL (1994) A histone-binding protein, nucleoplasmin, stimulates transcription factor binding to nucleosomes and factor-induced nucleosome disassembly. EMBO J 13:380–390
Cotten M, Chalkley R (1987) Purification of a novel, nucleoplasmin-like protein from somatic nuclei. EMBO J 6:3945–3954
Dilworth SM, Black SJ, Laskey RA (1987) Two complexes that contain histones are required for nucleosome assembly in vitro: role of nucleoplasmin and N1 in Xenopus egg extracts. Cell 51:1009–1018
Dingwall C, Dilworth SM, Black S, Kearsey SE, Cox LS, Laskey RA (1987) Nucleoplasmin cDNA sequence reveals polyglutamic acid tracts and a cluster of sequences homologous to putative nuclear localization signals. EMBO J 6:69–74
Dingwall C, Laskey RA (1991) Nuclear targeting sequences—a consensus? Trends Biochem Sci 16:478–481
Elgin SCR (1988) The formation and function of DNase I hypersensitive sites in the process of gene activation. J Biol Chem 263:19259–19262
Felsenfeld G (1992) Chromatin as an essential part of the transcriptional mechanism. Nature 355:219–223
Fotedar R, Roberts JM (1989) Multiple pathway for replication-dependent nucleosome assembly. Proc Natl Acad Sci USA 86:6459–6463
Fujii-Nakada T, Ishimi Y, Okuda A, Kikuchi A (1992) Functional analysis of nucleosome assembly protein, NAP-1. The negatively charged COOH-terminal regions is not necessary for the intrinsic assembly activity. J Biol Chem 267:20980–20986
Garcia-Bustos J, Heitman J, Hall MN (1991) Nuclear protein localization. Biochem Biophys Acta 1071:83–101
Gruss C, Gutierrez C, Burhans WC, DePamphilis ML, Koller T, Sogo JM (1990) Nucleosome assembly in mammalian cell extracts before and after DNA replication. EMBO J 9:2911–2922
Gruss C, Wu J, Koller T, Sogo JM (1993) Disruption of the nucleosome at the replication fork. EMBO J 12:4533–4545
Harlow E, Lane D (1988) Antibodies. Cold Spring Harbor Laboratory Press. Cold Spring Harbor, New York
Ishimi Y, Hirosumi J, Sato W, Sugasawa K, Yokota S, Hanaoka F, Yamada M (1984) Purification and initial characterization of a protein which facilitates assembly of nucleosome-like structure from mammalian cells. Eur J Biochem 142:431–439
Ishimi Y, Sato W, Kojima M, Sugasawa K, Hanaoka F, Yamada M (1985) Rapid purification of nucleosome assembly protein (AP-I) and production of monoclonal antibodies against it. Cell Struc Funct 10:373–382
Ishimi Y, Kojima M, Yamada M, Hanaoka F (1987) Binding mode of nucleosome-assembly protein (AP-I) and histones. Eur J Biochem 162:19–24
Ishimi Y, Claude A, Bullock P, Hurwitz J (1988) Complete enzymatic synthesis of DNA containing the SV 40 origin of replication. J Biol Chem 263:19723–19733
Ishimi Y, Kikuchi A (1991) Identification and molecular cloning of yeast homolog of nucleosome assembly protein I which facilitates nucleosome assembly in vitro. J Biol Chem 266:7025–7029
Jantzen HM, Admon A, Bell SP, Tjian R (1990) Nuclear transcription factor hUBF contains a DNA-binding motif with homology to HMG proteins. Nature 344:830–836
Jantzen HM, Chow AM, King DS, Tjian R (1992) Multiple domains of the RNA polymerase I activator hUBF interact with the TATA-binding protein complex hSL1 to mediate transcription. Genes Dev 6:1950–1963
Kalderon D, Roberts BL, Richardson WD, Smith AE (1984) A short amino acid sequence able to specify nuclear location. Cell 39:499–509
Kleinschmidt JA, Franke WW (1982) Soluble acidic complexes containing histones H3 and H4 in nuclei of Xenopus laevis oocytes. Cell 29:799–809
Kleinschmidt JA, Fortkamp E, Krohne G, Zentgraf H, Franke WW (1985) Co-existence of two different types of soluble histone complexes in nuclei of Xenopus laevis oocytes. EMBO J 260:1166–1176
Kleinschmidt JA, Dingwall C, Maier G, Franke WW (1986) Molecular characterization of a karyophilic, histone binding protein: cDNA cloning, amino acid sequence and expression of nuclear protein N1/N2 of Xenopus laevis. EMBO J 5:3547–3552
Kleinschmidt JA, Seiter A (1988) Identification of domains involved in nuclear uptake and histone binding of protein N1 of Xenopus laevis. EMBO J 7:1065–1614
Kleinschmidt JA, Seiter A, Zentgraf H (1990) Nucleosome assembly in vitro: separate histone transfer and synergistic interaction of native histone complexes purified from nuclei of Xenopus laevis oocytes. EMBO J 9:1309–1318
Kornberg RD, Lorch Y (1991) Irresistible force meets immovable object: transcription and nucleosome. Cell 67:833–836
Krude T, Knippers R (1991) Transfer of nucleosome from parental to replicated chromatin. Mol Cell Biol 11:6257–6267
Kyte J, Doolittle RF (1982) A simple method for displaying the hydropathic character of a protein. J Mol Biol 157:105–132
Lapeyre B, Bourbon H, Amalric F (1987) Nucleolin, the major nucleolar protein of growing eukaryotic cells: an unusual protein structure revealed by the nucleotide sequence. Proc Natl Acad Sci USA 84:1472–1476
Laskey RA, Honda BM, Mills AD, Finch JT (1978) Nucleosomes are assembled by an acidic protein which binds histones and transfers them to DNA. Nature 275:416–420
Lässle M, Richter A, Knippers R (1992) Comparison of replicative and non-replicative chromatin assembly pathways in HeLa cell extracts. Biochem Biophy Acta 1132:1–10
Lee DY, Hayes JJ, Pruss D, Wolffe AP (1993) A positive role for histone acetylation in transcription factor access to nucleosomal DNA. Cell 72:73–84
Lin K, Cheng S (1990) An efficient method to purify active eukaryotic proteins from the inclusion bodies in Escherichia coli. Biotechniques 11:748–752
Robbins J, Dilworth SM, Laskey RA, Dingwall C (1991) Two interdependent basic domains in nucleoplasmin nuclear targeting sequence: identification of a class of bipartite nuclear targeting sequence. Cell 64:615–623
Rodriguez-Campos A, Shimamura A, Worcel A (1989) Assembly and properties of chromatin containing histone H1. J Mol Biol 209:135–150
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci 74:5463–5467
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning. Cold Spring Harbor Laboratory Press. Cold Spring Harbor
Sapp M, Worcel A (1990) Purification and mechanism of action of a nucleosome assembly factor from Xenopus oocytes. J Biol Chem 265:9357–9365
Schmidt-Zachmann MS, Hugel-Dorr B, Franke WW (1987) A constitutive nucleolar protein identified as a member of the nucleoplasmin family. EMBO J 6:1881–1890
Shirakawa H, Tsuda K-I, Yoshida M (1990) Primary structure of non-histone chromosomal protein HMG2 revealed by the nucleotide sequence. Biochemistry 29:4419–4423
Simon HU, Millis GB, Kzlowsky M, Hogg D, Branch D, Ishimi Y, Siminovitch KA (1994) Molecular characterization of hNRP, a cDNA encoding a human nucleosome assembly protein Irelated gene product involved in the induction of cell proliferation. Biochem J 297:389–397
Smith S, Stillman B (1989) Purification and characterization of CAF-1, a human cell factor required for chromatin assembly during DNA replication in vitro. Cell 58:15–25
Smith S, Stillman B (1991) Stepwise assembly of chromatin during DNA replication in vitro. EMBO J 10:971–980
Studier FW, Rosenberg AH, Dunn JJ, Dubendorff JN (1990) Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol 185:60–89
Towbin HT, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354
Van Holde KE (1988) Chromatin. Springer-Verlag, New York
Workman JL, Buchman AR (1993) Multiple functions of nucleosome and regulatory factors in transcription. Trends Biochem Sci 18:90–95
Zucker K, Worcel A (1990) The histone H3/H4.N1 complex supplemented with histone H2A-H2B dimers and DNA topoisomerase I forms nucleosomes on circular DNA under physiological conditions. J Biol Chem 265:14487–14496
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Yoon, H.W., Kim, M.C., Lee, S.Y. et al. Molecular cloning and functional characterization of a cDNA encoding nucleosome assembly protein 1 (NAP-1) from soybean. Molec. Gen. Genet. 249, 465–473 (1995). https://doi.org/10.1007/BF00290572
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DOI: https://doi.org/10.1007/BF00290572