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. 1988 Dec 9;16(23):11107–11123. doi: 10.1093/nar/16.23.11107

Effects of high mobility group proteins 1 and 2 on initiation and elongation of specific transcription by RNA polymerase II in vitro.

D J Tremethick 1, P L Molloy 1
PMCID: PMC338999  PMID: 2462724

Abstract

High mobility group proteins 1 and 2 (HMGs 1 and 2) are abundant chromosomal proteins of higher eukaryotes, which have been found to be enriched in regions of active chromatin. We have previously demonstrated that they can stimulate specific transcription in vitro by RNA polymerases II and III and overcome inhibition caused by added histones. Here we study whether these effects are mediated at the level of initiation or elongation of transcription. Additions of HMGs 1 and 2 and/or histones were found to have only small or no effect on the efficiency of elongation; this was determined by comparing the relative synthesis of transcripts of different lengths, ranging from 95 to 1535 bases. The observed stimulation cannot be explained by an increased utilization of initiation complexes for multiple rounds of transcription as a similar level of stimulation by HMGs 1 and 2 was seen when RNA synthesis was limited to one round per template DNA by addition of a low level of Sarkosyl after formation of initiation complexes. The effects of HMGs 1 and 2 were principally seen on the rate of formation of effective initiation complexes. These data are consistent with the hypothesis that HMGs 1 and 2 stimulate transcription by facilitating the formation of active initiation complexes on template DNA.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ackerman S., Bunick D., Zandomeni R., Weinmann R. RNA polymerase II ternary transcription complexes generated in vitro. Nucleic Acids Res. 1983 Sep 10;11(17):6041–6064. doi: 10.1093/nar/11.17.6041. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Benezra R., Cantor C. R., Axel R. Nucleosomes are phased along the mouse beta-major globin gene in erythroid and nonerythroid cells. Cell. 1986 Mar 14;44(5):697–704. doi: 10.1016/0092-8674(86)90835-4. [DOI] [PubMed] [Google Scholar]
  3. Burton Z. F., Ortolan L. G., Greenblatt J. Proteins that bind to RNA polymerase II are required for accurate initiation of transcription at the adenovirus 2 major late promoter. EMBO J. 1986 Nov;5(11):2923–2930. doi: 10.1002/j.1460-2075.1986.tb04588.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Carthew R. W., Chodosh L. A., Sharp P. A. An RNA polymerase II transcription factor binds to an upstream element in the adenovirus major late promoter. Cell. 1985 Dec;43(2 Pt 1):439–448. doi: 10.1016/0092-8674(85)90174-6. [DOI] [PubMed] [Google Scholar]
  5. Cartwright I. L., Elgin S. C. Nucleosomal instability and induction of new upstream protein-DNA associations accompany activation of four small heat shock protein genes in Drosophila melanogaster. Mol Cell Biol. 1986 Mar;6(3):779–791. doi: 10.1128/mcb.6.3.779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chambers S. A., Rill R. L. Enrichment of transcribed and newly replicated DNA in soluble chromatin released from nuclei by mild micrococcal nuclease digestion. Biochim Biophys Acta. 1984 Jun 16;782(2):202–209. doi: 10.1016/0167-4781(84)90025-3. [DOI] [PubMed] [Google Scholar]
  7. Chrysogelos S., Riley D. E., Stein G., Stein J. A human histone H4 gene exhibits cell cycle-dependent changes in chromatin structure that correlate with its expression. Proc Natl Acad Sci U S A. 1985 Nov;82(22):7535–7539. doi: 10.1073/pnas.82.22.7535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cohen R. B., Sheffery M. Nucleosome disruption precedes transcription and is largely limited to the transcribed domain of globin genes in murine erythroleukemia cells. J Mol Biol. 1985 Mar 5;182(1):109–129. doi: 10.1016/0022-2836(85)90031-2. [DOI] [PubMed] [Google Scholar]
  9. Culotta V. C., Wides R. J., Sollner-Webb B. Eucaryotic transcription complexes are specifically associated in large sedimentable structures: rapid isolation of polymerase I, II, and III transcription factors. Mol Cell Biol. 1985 Jul;5(7):1582–1590. doi: 10.1128/mcb.5.7.1582. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Davison B. L., Egly J. M., Mulvihill E. R., Chambon P. Formation of stable preinitiation complexes between eukaryotic class B transcription factors and promoter sequences. Nature. 1983 Feb 24;301(5902):680–686. doi: 10.1038/301680a0. [DOI] [PubMed] [Google Scholar]
  11. Eissenberg J. C., Cartwright I. L., Thomas G. H., Elgin S. C. Selected topics in chromatin structure. Annu Rev Genet. 1985;19:485–536. doi: 10.1146/annurev.ge.19.120185.002413. [DOI] [PubMed] [Google Scholar]
  12. Fire A., Samuels M., Sharp P. A. Interactions between RNA polymerase II, factors, and template leading to accurate transcription. J Biol Chem. 1984 Feb 25;259(4):2509–2516. [PubMed] [Google Scholar]
  13. Gill G., Ptashne M. Mutants of GAL4 protein altered in an activation function. Cell. 1987 Oct 9;51(1):121–126. doi: 10.1016/0092-8674(87)90016-x. [DOI] [PubMed] [Google Scholar]
  14. Hawley D. K., Roeder R. G. Separation and partial characterization of three functional steps in transcription initiation by human RNA polymerase II. J Biol Chem. 1985 Jul 5;260(13):8163–8172. [PubMed] [Google Scholar]
  15. Hope I. A., Struhl K. Functional dissection of a eukaryotic transcriptional activator protein, GCN4 of yeast. Cell. 1986 Sep 12;46(6):885–894. doi: 10.1016/0092-8674(86)90070-x. [DOI] [PubMed] [Google Scholar]
  16. Hough P. V., Mastrangelo I. A., Wall J. S., Hainfeld J. F., Simon M. N., Manley J. L. DNA-protein complexes spread on N2-discharged carbon film and characterized by molecular weight and its projected distribution. J Mol Biol. 1982 Sep 15;160(2):375–386. doi: 10.1016/0022-2836(82)90183-8. [DOI] [PubMed] [Google Scholar]
  17. Jackson J. B., Pollock J. M., Jr, Rill R. L. Chromatin fractionation procedure that yields nucleosomes containing near-stoichiometric amounts of high mobility group nonhistone chromosomal proteins. Biochemistry. 1979 Aug 21;18(17):3739–3748. doi: 10.1021/bi00584a015. [DOI] [PubMed] [Google Scholar]
  18. Jackson J. B., Rill R. L. Circular dichroism, thermal denaturation, and deoxyribonuclease I digestion studies of nucleosomes highly enriched in high mobility group proteins HMG 1 and HMG 2. Biochemistry. 1981 Feb 17;20(4):1042–1046. doi: 10.1021/bi00507a060. [DOI] [PubMed] [Google Scholar]
  19. Knezetic J. A., Luse D. S. The presence of nucleosomes on a DNA template prevents initiation by RNA polymerase II in vitro. Cell. 1986 Apr 11;45(1):95–104. doi: 10.1016/0092-8674(86)90541-6. [DOI] [PubMed] [Google Scholar]
  20. Kuehl L., Lyness T., Dixon G. H., Levy-Wilson B. Distribution of high mobility group proteins among domains of trout testis chromatin differing in their susceptibility to micrococcal nuclease. J Biol Chem. 1980 Feb 10;255(3):1090–1095. [PubMed] [Google Scholar]
  21. Lorch Y., LaPointe J. W., Kornberg R. D. Nucleosomes inhibit the initiation of transcription but allow chain elongation with the displacement of histones. Cell. 1987 Apr 24;49(2):203–210. doi: 10.1016/0092-8674(87)90561-7. [DOI] [PubMed] [Google Scholar]
  22. Losa R., Brown D. D. A bacteriophage RNA polymerase transcribes in vitro through a nucleosome core without displacing it. Cell. 1987 Aug 28;50(5):801–808. doi: 10.1016/0092-8674(87)90338-2. [DOI] [PubMed] [Google Scholar]
  23. Luse D. S., Kochel T., Kuempel E. D., Coppola J. A., Cai H. Transcription initiation by RNA polymerase II in vitro. At least two nucleotides must be added to form a stable ternary complex. J Biol Chem. 1987 Jan 5;262(1):289–297. [PubMed] [Google Scholar]
  24. Manley J. L., Colozzo M. T. Synthesis in vitro of an exceptionally long RNA transcript promoted by an AluI sequence. Nature. 1982 Nov 25;300(5890):376–379. doi: 10.1038/300376a0. [DOI] [PubMed] [Google Scholar]
  25. Manley J. L., Fire A., Cano A., Sharp P. A., Gefter M. L. DNA-dependent transcription of adenovirus genes in a soluble whole-cell extract. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3855–3859. doi: 10.1073/pnas.77.7.3855. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Matsui T. Transcription of adenovirus 2 major late and peptide IX genes under conditions of in vitro nucleosome assembly. Mol Cell Biol. 1987 Apr;7(4):1401–1408. doi: 10.1128/mcb.7.4.1401. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Moncollin V., Miyamoto N. G., Zheng X. M., Egly J. M. Purification of a factor specific for the upstream element of the adenovirus-2 major late promoter. EMBO J. 1986 Oct;5(10):2577–2584. doi: 10.1002/j.1460-2075.1986.tb04537.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Moreno M. L., Chrysogelos S. A., Stein G. S., Stein J. L. Reversible changes in the nucleosomal organization of a human H4 histone gene during the cell cycle. Biochemistry. 1986 Sep 23;25(19):5364–5370. doi: 10.1021/bi00367a003. [DOI] [PubMed] [Google Scholar]
  29. Reeves R. Transcriptionally active chromatin. Biochim Biophys Acta. 1984 Sep 10;782(4):343–393. doi: 10.1016/0167-4781(84)90044-7. [DOI] [PubMed] [Google Scholar]
  30. Reinberg D., Horikoshi M., Roeder R. G. Factors involved in specific transcription in mammalian RNA polymerase II. Functional analysis of initiation factors IIA and IID and identification of a new factor operating at sequences downstream of the initiation site. J Biol Chem. 1987 Mar 5;262(7):3322–3330. [PubMed] [Google Scholar]
  31. Reinberg D., Roeder R. G. Factors involved in specific transcription by mammalian RNA polymerase II. Purification and functional analysis of initiation factors IIB and IIE. J Biol Chem. 1987 Mar 5;262(7):3310–3321. [PubMed] [Google Scholar]
  32. Rose S. M., Garrard W. T. Differentiation-dependent chromatin alterations precede and accompany transcription of immunoglobulin light chain genes. J Biol Chem. 1984 Jul 10;259(13):8534–8544. [PubMed] [Google Scholar]
  33. Safer B., Yang L., Tolunay H. E., Anderson W. F. Isolation of stable preinitiation, initiation, and elongation complexes from RNA polymerase II-directed transcription. Proc Natl Acad Sci U S A. 1985 May;82(9):2632–2636. doi: 10.1073/pnas.82.9.2632. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Sawadogo M., Roeder R. G. Energy requirement for specific transcription initiation by the human RNA polymerase II system. J Biol Chem. 1984 Apr 25;259(8):5321–5326. [PubMed] [Google Scholar]
  35. Sawadogo M., Roeder R. G. Factors involved in specific transcription by human RNA polymerase II: analysis by a rapid and quantitative in vitro assay. Proc Natl Acad Sci U S A. 1985 Jul;82(13):4394–4398. doi: 10.1073/pnas.82.13.4394. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Sawadogo M., Roeder R. G. Interaction of a gene-specific transcription factor with the adenovirus major late promoter upstream of the TATA box region. Cell. 1985 Nov;43(1):165–175. doi: 10.1016/0092-8674(85)90021-2. [DOI] [PubMed] [Google Scholar]
  37. Sergeant A., Bohmann D., Zentgraf H., Weiher H., Keller W. A transcription enhancer acts in vitro over distances of hundreds of base-pairs on both circular and linear templates but not on chromatin-reconstituted DNA. J Mol Biol. 1984 Dec 15;180(3):577–600. doi: 10.1016/0022-2836(84)90028-7. [DOI] [PubMed] [Google Scholar]
  38. Sun Y. L., Xu Y. Z., Bellard M., Chambon P. Digestion of the chicken beta-globin gene chromatin with micrococcal nuclease reveals the presence of an altered nucleosomal array characterized by an atypical ladder of DNA fragments. EMBO J. 1986 Feb;5(2):293–300. doi: 10.1002/j.1460-2075.1986.tb04212.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Tolunay H. E., Yang L., Anderson W. F., Safer B. Isolation of an active transcription initiation complex from HeLa cell-free extract. Proc Natl Acad Sci U S A. 1984 Oct;81(19):5916–5920. doi: 10.1073/pnas.81.19.5916. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Tremethick D. J., Molloy P. L. High mobility group proteins 1 and 2 stimulate transcription in vitro by RNA polymerases II and III. J Biol Chem. 1986 May 25;261(15):6986–6992. [PubMed] [Google Scholar]
  41. Tsuda M., Hirose S., Suzuki Y. Participation of the upstream region of the fibroin gene in the formation of transcription complex in vitro. Mol Cell Biol. 1986 Nov;6(11):3928–3933. doi: 10.1128/mcb.6.11.3928. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Watt F., Molloy P. L. High mobility group proteins 1 and 2 stimulate binding of a specific transcription factor to the adenovirus major late promoter. Nucleic Acids Res. 1988 Feb 25;16(4):1471–1486. doi: 10.1093/nar/16.4.1471. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Weintraub H. Assembly and propagation of repressed and depressed chromosomal states. Cell. 1985 Oct;42(3):705–711. doi: 10.1016/0092-8674(85)90267-3. [DOI] [PubMed] [Google Scholar]
  44. Workman J. L., Roeder R. G. Binding of transcription factor TFIID to the major late promoter during in vitro nucleosome assembly potentiates subsequent initiation by RNA polymerase II. Cell. 1987 Nov 20;51(4):613–622. doi: 10.1016/0092-8674(87)90130-9. [DOI] [PubMed] [Google Scholar]
  45. Yu J., Smith R. D. Sequential alterations in globin gene chromatin structure during erythroleukemia cell differentiation. J Biol Chem. 1985 Mar 10;260(5):3035–3040. [PubMed] [Google Scholar]
  46. Zheng X. M., Moncollin V., Egly J. M., Chambon P. A general transcription factor forms a stable complex with RNA polymerase B (II). Cell. 1987 Jul 31;50(3):361–368. doi: 10.1016/0092-8674(87)90490-9. [DOI] [PubMed] [Google Scholar]
  47. ten Heggeler B., Wahli W. Visualization of RNA polymerase II ternary transcription complexes formed in vitro on a Xenopus laevis vitellogenin gene. EMBO J. 1985 Sep;4(9):2269–2273. doi: 10.1002/j.1460-2075.1985.tb03925.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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