PMCCPMCCPMCC

Search tips
Search criteria 

Advanced

 
Logo of molcellbPermissionsJournals.ASM.orgJournalMCB ArticleJournal InfoAuthorsReviewers
 
Mol Cell Biol. 1987 November; 7(11): 4048–4057.
PMCID: PMC368075

Amino acid sequences that determine the nuclear localization of yeast histone 2B.

Abstract

Histone-beta-galactosidase protein fusions were used to identify the domain of yeast histone 2B, which targets this protein to the nucleus. Amino acids 28 to 33 in H2B were required for nuclear localization of such fusion proteins and thus constitute a nuclear localization sequence. The amino acid sequence in this region (Gly-29 Lys Lys Arg Ser Lys Ala) is similar to the nuclear location signal in simian virus 40 large T antigen (Pro-126 Lys Lys Lys Arg Lys Val) (D. Kalderon, B.L. Roberts, W.D. Richardson, and A.E. Smith, Cell 39:499-509, 1984). A point mutation changing lysine 31 to methionine abolished nuclear localization of an H2B-beta-galactosidase fusion protein containing amino acids 1 to 33 of H2B. However, an H2B-beta-galactosidase fusion protein containing both this point mutation and the H2A interaction domain of H2B was nuclear localized. These results suggest that H2A and H2B may be cotransported to the nucleus as a heterodimer.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (2.6M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Adams AE, Pringle JR. Relationship of actin and tubulin distribution to bud growth in wild-type and morphogenetic-mutant Saccharomyces cerevisiae. J Cell Biol. 1984 Mar;98(3):934–945. [PMC free article] [PubMed]
  • Bendayan M, Zollinger M. Ultrastructural localization of antigenic sites on osmium-fixed tissues applying the protein A-gold technique. J Histochem Cytochem. 1983 Jan;31(1):101–109. [PubMed]
  • Blake MS, Johnston KH, Russell-Jones GJ, Gotschlich EC. A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on Western blots. Anal Biochem. 1984 Jan;136(1):175–179. [PubMed]
  • Bonner WM. Protein migration into nuclei. I. Frog oocyte nuclei in vivo accumulate microinjected histones, allow entry to small proteins, and exclude large proteins. J Cell Biol. 1975 Feb;64(2):421–430. [PMC free article] [PubMed]
  • Bonner WM. Protein migration into nuclei. II. Frog oocyte nuclei accumulate a class of microinjected oocyte nuclear proteins and exclude a class of microinjected oocyte cytoplasmic proteins. J Cell Biol. 1975 Feb;64(2):431–437. [PMC free article] [PubMed]
  • Botstein D, Falco SC, Stewart SE, Brennan M, Scherer S, Stinchcomb DT, Struhl K, Davis RW. Sterile host yeasts (SHY): a eukaryotic system of biological containment for recombinant DNA experiments. Gene. 1979 Dec;8(1):17–24. [PubMed]
  • Chou PY, Fasman GD. Empirical predictions of protein conformation. Annu Rev Biochem. 1978;47:251–276. [PubMed]
  • Davey J, Dimmock NJ, Colman A. Identification of the sequence responsible for the nuclear accumulation of the influenza virus nucleoprotein in Xenopus oocytes. Cell. 1985 Mar;40(3):667–675. [PubMed]
  • De Robertis EM, Longthorne RF, Gurdon JB. Intracellular migration of nuclear proteins in Xenopus oocytes. Nature. 1978 Mar 16;272(5650):254–256. [PubMed]
  • Dingwall C, Sharnick SV, Laskey RA. A polypeptide domain that specifies migration of nucleoplasmin into the nucleus. Cell. 1982 Sep;30(2):449–458. [PubMed]
  • Feldherr CM, Kallenbach E, Schultz N. Movement of a karyophilic protein through the nuclear pores of oocytes. J Cell Biol. 1984 Dec;99(6):2216–2222. [PMC free article] [PubMed]
  • Fried HM, Nam HG, Loechel S, Teem J. Characterization of yeast strains with conditionally expressed variants of ribosomal protein genes tcm1 and cyh2. Mol Cell Biol. 1985 Jan;5(1):99–108. [PMC free article] [PubMed]
  • Gillam S, Astell CR, Smith M. Site-specific mutagenesis using oligodeoxyribonucleotides: isolation of a phenotypically silent phi X174 mutant, with a specific nucleotide deletion, at very high efficiency. Gene. 1980 Dec;12(1-2):129–137. [PubMed]
  • Gosselin EJ, Sorenson GD, Dennett JC, Cate CC. Unlabeled antibody methods in electron microscopy: a comparison of single and multistep procedures using colloidal gold. J Histochem Cytochem. 1984 Aug;32(8):799–804. [PubMed]
  • Guarente L, Lauer G, Roberts TM, Ptashne M. Improved methods for maximizing expression of a cloned gene: a bacterium that synthesizes rabbit beta-globin. Cell. 1980 Jun;20(2):543–553. [PubMed]
  • Guarente L, Yocum RR, Gifford P. A GAL10-CYC1 hybrid yeast promoter identifies the GAL4 regulatory region as an upstream site. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7410–7414. [PubMed]
  • Hall MN, Hereford L, Herskowitz I. Targeting of E. coli beta-galactosidase to the nucleus in yeast. Cell. 1984 Apr;36(4):1057–1065. [PubMed]
  • Hereford L, Bromley S, Osley MA. Periodic transcription of yeast histone genes. Cell. 1982 Aug;30(1):305–310. [PubMed]
  • Hereford L, Fahrner K, Woolford J, Jr, Rosbash M, Kaback DB. Isolation of yeast histone genes H2A and H2B. Cell. 1979 Dec;18(4):1261–1271. [PubMed]
  • Hereford LM, Osley MA, Ludwig TR, 2nd, McLaughlin CS. Cell-cycle regulation of yeast histone mRNA. Cell. 1981 May;24(2):367–375. [PubMed]
  • Isenberg I. Histones. Annu Rev Biochem. 1979;48:159–191. [PubMed]
  • Ito H, Fukuda Y, Murata K, Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. [PMC free article] [PubMed]
  • Kalderon D, Richardson WD, Markham AF, Smith AE. Sequence requirements for nuclear location of simian virus 40 large-T antigen. Nature. 1984 Sep 6;311(5981):33–38. [PubMed]
  • Kalderon D, Roberts BL, Richardson WD, Smith AE. A short amino acid sequence able to specify nuclear location. Cell. 1984 Dec;39(3 Pt 2):499–509. [PubMed]
  • Kyte J, Doolittle RF. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. [PubMed]
  • Lanford RE, Butel JS. Construction and characterization of an SV40 mutant defective in nuclear transport of T antigen. Cell. 1984 Jul;37(3):801–813. [PubMed]
  • Lanford RE, Kanda P, Kennedy RC. Induction of nuclear transport with a synthetic peptide homologous to the SV40 T antigen transport signal. Cell. 1986 Aug 15;46(4):575–582. [PubMed]
  • Leary JJ, Brigati DJ, Ward DC. Rapid and sensitive colorimetric method for visualizing biotin-labeled DNA probes hybridized to DNA or RNA immobilized on nitrocellulose: Bio-blots. Proc Natl Acad Sci U S A. 1983 Jul;80(13):4045–4049. [PubMed]
  • Mardian JK, Isenberg I. Yeast inner histones and the evolutionary conservation of histone-histone interactions. Biochemistry. 1978 Sep 5;17(18):3825–3833. [PubMed]
  • McGhee JD, Felsenfeld G. Nucleosome structure. Annu Rev Biochem. 1980;49:1115–1156. [PubMed]
  • Melton DA, Krieg PA, Rebagliati MR, Maniatis T, Zinn K, Green MR. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. [PMC free article] [PubMed]
  • Messing J, Crea R, Seeburg PH. A system for shotgun DNA sequencing. Nucleic Acids Res. 1981 Jan 24;9(2):309–321. [PMC free article] [PubMed]
  • Moreland RB, Nam HG, Hereford LM, Fried HM. Identification of a nuclear localization signal of a yeast ribosomal protein. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6561–6565. [PubMed]
  • Moss T, Cary PD, Abercrombie BD, Crane-Robinson C, Bradbury EM. A pH-dependent interaction between histones H2A and H2B involving secondary and tertiary folding. Eur J Biochem. 1976 Dec 11;71(2):337–350. [PubMed]
  • Osley MA, Gould J, Kim S, Kane MY, Hereford L. Identification of sequences in a yeast histone promoter involved in periodic transcription. Cell. 1986 May 23;45(4):537–544. [PubMed]
  • Osley MA, Hereford L. Identification of a sequence responsible for periodic synthesis of yeast histone 2A mRNA. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7689–7693. [PubMed]
  • Paine PL, Moore LC, Horowitz SB. Nuclear envelope permeability. Nature. 1975 Mar 13;254(5496):109–114. [PubMed]
  • Paucha E, Kalderon D, Richardson WD, Harvey RW, Smith AE. The abnormal location of cytoplasmic SV40 large T is not caused by failure to bind to DNA or to p53. EMBO J. 1985 Dec 1;4(12):3235–3240. [PubMed]
  • Peters R. Nucleo-cytoplasmic flux and intracellular mobility in single hepatocytes measured by fluorescence microphotolysis. EMBO J. 1984 Aug;3(8):1831–1836. [PubMed]
  • Pustell J, Kafatos FC. A convenient and adaptable package of computer programs for DNA and protein sequence management, analysis and homology determination. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 2):643–655. [PMC free article] [PubMed]
  • Richardson WD, Roberts BL, Smith AE. Nuclear location signals in polyoma virus large-T. Cell. 1986 Jan 17;44(1):77–85. [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PubMed]
  • Schuster T, Han M, Grunstein M. Yeast histone H2A and H2B amino termini have interchangeable functions. Cell. 1986 May 9;45(3):445–451. [PubMed]
  • Silver PA, Brent R, Ptashne M. DNA binding is not sufficient for nuclear localization of regulatory proteins in Saccharomyces cerevisiae. Mol Cell Biol. 1986 Dec;6(12):4763–4766. [PMC free article] [PubMed]
  • Silver PA, Keegan LP, Ptashne M. Amino terminus of the yeast GAL4 gene product is sufficient for nuclear localization. Proc Natl Acad Sci U S A. 1984 Oct;81(19):5951–5955. [PubMed]
  • Smith AE, Kalderon D, Roberts BL, Colledge WH, Edge M, Gillett P, Markham A, Paucha E, Richardson WD. The nuclear location signal. Proc R Soc Lond B Biol Sci. 1985 Oct 22;226(1242):43–58. [PubMed]
  • Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. [PubMed]
  • Wallis JW, Hereford L, Grunstein M. Histone H2B genes of yeast encode two different proteins. Cell. 1980 Dec;22(3):799–805. [PubMed]
  • Wallis JW, Rykowski M, Grunstein M. Yeast histone H2B containing large amino terminus deletions can function in vivo. Cell. 1983 Dec;35(3 Pt 2):711–719. [PubMed]

Articles from Molecular and Cellular Biology are provided here courtesy of American Society for Microbiology (ASM)