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PMID: 11040216 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Spt5 and spt6 are associated with active transcription and have characteristics of general elongation factors in D. melanogaster.

Genes & development ·Vol. 14 ·No. 20 ·2000-10-15 ·Pages 2623-34

Kaplan CD, Morris JR, Wu C, Winston F

Abstract

The Spt4, Spt5, and Spt6 proteins are conserved throughout eukaryotes and are believed to play critical and related roles in transcription. They have a positive role in transcription elongation in Saccharomyces cerevisiae and in the activation of transcription by the HIV Tat protein in human cells. In contrast, a complex of Spt4 and Spt5 is required in vitro for the inhibition of RNA polymerase II (Pol II) elongation by the drug DRB, suggesting also a negative role in vivo. To learn more about the function of the Spt4/Spt5 complex and Spt6 in vivo, we have identified Drosophila homologs of Spt5 and Spt6 and characterized their localization on Drosophila polytene chromosomes. We find that Spt5 and Spt6 localize extensively with the phosphorylated, actively elongating form of Pol II, to transcriptionally active sites during salivary gland development and upon heat shock. Furthermore, Spt5 and Spt6 do not colocalize widely with the unphosphorylated, nonelongating form of Pol II. These results strongly suggest that Spt5 and Spt6 play closely related roles associated with active transcription in vivo.

MeSH Terms
Animals Chromosomal Proteins, Non-Histone Chromosomes/metabolism Cyclin T Cyclins/metabolism Drosophila Proteins Drosophila melanogaster/embryology,genetics Embryo, Nonmammalian Fungal Proteins/genetics,metabolism Gene Expression Regulation, Developmental Glue Proteins, Drosophila/genetics,metabolism Heat-Shock Response Histone Chaperones Insect Proteins/genetics,metabolism Molecular Sequence Data Nuclear Proteins/genetics,metabolism Peptide Elongation Factors/genetics,metabolism RNA Polymerase II/genetics,metabolism Saccharomyces cerevisiae Proteins Sequence Analysis Transcription, Genetic Transcriptional Elongation Factors
Chemicals
CCNT1 protein, human Chromosomal Proteins, Non-Histone CycT protein, Drosophila Cyclin T Cyclins Drosophila Proteins Fungal Proteins Glue Proteins, Drosophila Histone Chaperones Insect Proteins Nuclear Proteins Peptide Elongation Factors SPT6 protein, S cerevisiae Saccharomyces cerevisiae Proteins Transcriptional Elongation Factors SPT5 transcriptional elongation factor RNA Polymerase II
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Kaplan C D
Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA.
Morris J R
Wu C
Winston F
References (83)
83 references, click to expand
  1. The helix-hairpin-helix DNA-binding motif: a structural basis for non-sequence-specific recognition of DNA.
    Nucleic Acids Res. 1996 Jul 1;24(13):2488-97 PMID: 8692686
  2. The C-terminal domain of the largest subunit of RNA polymerase II interacts with a novel set of serine/arginine-rich proteins.
    Proc Natl Acad Sci U S A. 1996 Jul 9;93(14):6975-80 PMID: 8692929
  3. Reversible phosphorylation of the C-terminal domain of RNA polymerase II.
    J Biol Chem. 1996 Aug 9;271(32):19009-12 PMID: 8759772
  4. A new gene locus of Bordetella pertussis defines a novel family of prokaryotic transcriptional accessory proteins.
    J Bacteriol. 1996 Aug;178(15):4445-52 PMID: 8755871
  5. Suppression analysis reveals a functional difference between the serines in positions two and five in the consensus sequence of the C-terminal domain of yeast RNA polymerase II.
    Genetics. 1996 Jun;143(2):661-71 PMID: 8725217
  6. HSF recruitment and loss at most Drosophila heat shock loci is coordinated and depends on proximal promoter sequences.
    Chromosoma. 1996 Sep;105(3):158-71 PMID: 8781184
  7. Mutations in the SPT4, SPT5, and SPT6 genes alter transcription of a subset of histone genes in Saccharomyces cerevisiae.
    Genetics. 1996 Aug;143(4):1543-54 PMID: 8844144
  8. P-TEFb, a cyclin-dependent kinase controlling elongation by RNA polymerase II.
    Mol Cell Biol. 2000 Apr;20(8):2629-34 PMID: 10733565
  9. Domains in the SPT5 protein that modulate its transcriptional regulatory properties.
    Mol Cell Biol. 2000 May;20(9):2970-83 PMID: 10757782
  10. P-TEFb kinase recruitment and function at heat shock loci.
    Genes Dev. 2000 Apr 1;14(7):792-803 PMID: 10766736
  11. FACT relieves DSIF/NELF-mediated inhibition of transcriptional elongation and reveals functional differences between P-TEFb and TFIIH.
    Mol Cell. 2000 Jun;5(6):1067-72 PMID: 10912001
  12. High-resolution localization of Drosophila Spt5 and Spt6 at heat shock genes in vivo: roles in promoter proximal pausing and transcription elongation.
    Genes Dev. 2000 Oct 15;14(20):2635-49 PMID: 11040217
  13. Patterns of puffing activity in the salivary gland chromosomes of Drosophila. I. Autosomal puffing patterns in a laboratory stock of Drosophila melanogaster.
    Chromosoma. 1967;21(4):398-428 PMID: 6056758
  14. Patterns of puffing activity in the salivary gland chromosomes of Drosophila. VI. Induction by ecdysone in salivary glands of D. melanogaster cultured in vitro.
    Chromosoma. 1972;38(3):255-81 PMID: 4627363
  15. Temporal control of puffing activity in polytene chromosomes.
    Cold Spring Harb Symp Quant Biol. 1974;38:655-62 PMID: 4208797
  16. Cloning and characterization of nine heat-shock-induced mRNAs of Drosophila melanogaster.
    Gene. 1981 Oct;15(1):67-80 PMID: 6795091
  17. A fluorescent quantitative PCR approach to map gene deletions in the Drosophila genome.
    Genetics. 1999 Nov;153(3):1313-6 PMID: 10545461
  18. Cytogenetic and genetic mapping of a salivary gland secretion protein in Drosophila melanogaster.
    Chromosoma. 1981;84(2):173-85 PMID: 6799259
  19. Pig heart calpastatin: identification of repetitive domain structures and anomalous behavior in polyacrylamide gel electrophoresis.
    Biochemistry. 1988 Mar 22;27(6):1964-72 PMID: 2837276
  20. Puffs and PCR: the in vivo dynamics of early gene expression during ecdysone responses in Drosophila.
    Development. 1993 Jun;118(2):613-27 PMID: 8223281
  21. Locus-specific variation in phosphorylation state of RNA polymerase II in vivo: correlations with gene activity and transcript processing.
    Genes Dev. 1993 Dec;7(12A):2329-44 PMID: 8253380
  22. Cytostellin distributes to nuclear regions enriched with splicing factors.
    J Cell Sci. 1994 Mar;107 ( Pt 3):387-96 PMID: 8006060
  23. Phosphorylation of RNA polymerase II C-terminal domain and transcriptional elongation.
    Nature. 1994 Jul 7;370(6484):75-7 PMID: 8015613
  24. Lentivirus Tat proteins specifically associate with a cellular protein kinase, TAK, that hyperphosphorylates the carboxyl-terminal domain of the large subunit of RNA polymerase II: candidate for a Tat cofactor.
    J Virol. 1995 Mar;69(3):1612-20 PMID: 7853496
  25. Imaging neuronal subsets and other cell types in whole-mount Drosophila embryos and larvae using antibody probes.
    Methods Cell Biol. 1994;44:445-87 PMID: 7707967
  26. Purification of P-TEFb, a transcription factor required for the transition into productive elongation.
    J Biol Chem. 1995 May 26;270(21):12335-8 PMID: 7759473
  27. Use of insoluble fusion proteins to purify antibodies.
    Biotechniques. 1995 Oct;19(4):544-6, 548, 550 PMID: 8777040
  28. Construction and analysis of yeast RNA polymerase II CTD deletion and substitution mutations.
    Genetics. 1995 Aug;140(4):1223-33 PMID: 7498765
  29. Drosophila Sgs genes: stage and tissue specificity of hormone responsiveness.
    Bioessays. 1996 Jan;18(1):47-54 PMID: 8593163
  30. Evidence that Spt6p controls chromatin structure by a direct interaction with histones.
    Science. 1996 Jun 7;272(5267):1473-6 PMID: 8633238
  31. Promoter is an important determinant of developmentally regulated puffing at the Sgs-4 locus of Drosophila melanogaster.
    Dev Biol. 1990 Apr;138(2):324-37 PMID: 2156737
  32. SPT6, an essential gene that affects transcription in Saccharomyces cerevisiae, encodes a nuclear protein with an extremely acidic amino terminus.
    Mol Cell Biol. 1990 Sep;10(9):4935-41 PMID: 2201908
  33. Basic local alignment search tool.
    J Mol Biol. 1990 Oct 5;215(3):403-10 PMID: 2231712
  34. Puffs and gene regulation--molecular insights into the Drosophila ecdysone regulatory hierarchy.
    Bioessays. 1990 Dec;12(12):561-8 PMID: 2127884
  35. SPT5, an essential gene important for normal transcription in Saccharomyces cerevisiae, encodes an acidic nuclear protein with a carboxy-terminal repeat.
    Mol Cell Biol. 1991 Jun;11(6):3009-19 PMID: 1840633
  36. Mutations in SPT16/CDC68 suppress cis- and trans-acting mutations that affect promoter function in Saccharomyces cerevisiae.
    Mol Cell Biol. 1991 Nov;11(11):5710-7 PMID: 1922073
  37. The RNA polymerase II general elongation factors.
    Trends Biochem Sci. 1996 Sep;21(9):351-5 PMID: 8870500
  38. The solution structure of the S1 RNA binding domain: a member of an ancient nucleic acid-binding fold.
    Cell. 1997 Jan 24;88(2):235-42 PMID: 9008164
  39. Splicing factors associate with hyperphosphorylated RNA polymerase II in the absence of pre-mRNA.
    J Cell Biol. 1997 Jan 13;136(1):19-28 PMID: 9008700
  40. Nucleosomes and transcription: recent lessons from genetics.
    Curr Opin Genet Dev. 1997 Apr;7(2):192-8 PMID: 9115423
  41. Human Supt5h protein, a putative modulator of chromatin structure, is reversibly phosphorylated in mitosis.
    FEBS Lett. 1997 Jun 2;409(1):74-8 PMID: 9199507
  42. The H3/H4 tetramer blocks transcript elongation by RNA polymerase II in vitro.
    J Biol Chem. 1997 Sep 12;272(37):23427-34 PMID: 9287358
  43. Taking a new TAK on tat transactivation.
    Genes Dev. 1997 Oct 15;11(20):2593-9 PMID: 9334323
  44. The Med proteins of yeast and their function through the RNA polymerase II carboxy-terminal domain.
    Genes Dev. 1998 Jan 1;12(1):45-54 PMID: 9420330
  45. DSIF, a novel transcription elongation factor that regulates RNA polymerase II processivity, is composed of human Spt4 and Spt5 homologs.
    Genes Dev. 1998 Feb 1;12(3):343-56 PMID: 9450929
  46. Evidence that Spt4, Spt5, and Spt6 control transcription elongation by RNA polymerase II in Saccharomyces cerevisiae.
    Genes Dev. 1998 Feb 1;12(3):357-69 PMID: 9450930
  47. Growth-related changes in phosphorylation of yeast RNA polymerase II.
    J Biol Chem. 1998 Feb 20;273(8):4689-94 PMID: 9468530
  48. The RNA polymerase II general elongation complex.
    Biol Chem. 1998 Jan;379(1):27-31 PMID: 9504713
  49. Role of the human homolog of the yeast transcription factor SPT5 in HIV-1 Tat-activation.
    J Mol Biol. 1998 Mar 27;277(2):179-97 PMID: 9514752
  50. Combinatorial effects of NusA and NusG on transcription elongation and Rho-dependent termination in Escherichia coli.
    J Mol Biol. 1998 May 1;278(2):307-16 PMID: 9571053
  51. Interplay between positive and negative elongation factors: drawing a new view of DRB.
    Genes Cells. 1998 Jan;3(1):9-15 PMID: 9581978
  52. FCP1, the RAP74-interacting subunit of a human protein phosphatase that dephosphorylates the carboxyl-terminal domain of RNA polymerase IIO.
    J Biol Chem. 1998 Oct 16;273(42):27593-601 PMID: 9765293
  53. Factors regulating the transcriptional elongation activity of RNA polymerase II.
    FASEB J. 1998 Nov;12(14):1437-46 PMID: 9806752
  54. The transcriptional switch between the Drosophila genes Pig-1 and Sgs-4 depends on a SEBP1 binding site within a shared enhancer region.
    Mol Gen Genet. 1998 Oct;259(6):656-63 PMID: 9819059
  55. Evidence that P-TEFb alleviates the negative effect of DSIF on RNA polymerase II-dependent transcription in vitro.
    EMBO J. 1998 Dec 15;17(24):7395-403 PMID: 9857195
  56. Conserved domains in DNA repair proteins and evolution of repair systems.
    Nucleic Acids Res. 1999 Mar 1;27(5):1223-42 PMID: 9973609
  57. Elongator, a multisubunit component of a novel RNA polymerase II holoenzyme for transcriptional elongation.
    Mol Cell. 1999 Jan;3(1):109-18 PMID: 10024884
  58. Structure and function of the human transcription elongation factor DSIF.
    J Biol Chem. 1999 Mar 19;274(12):8085-92 PMID: 10075709
  59. NELF, a multisubunit complex containing RD, cooperates with DSIF to repress RNA polymerase II elongation.
    Cell. 1999 Apr 2;97(1):41-51 PMID: 10199401
  60. The transition from initiation to elongation by RNA polymerase II.
    Cold Spring Harb Symp Quant Biol. 1998;63:289-300 PMID: 10384293
  61. Antitermination by bacteriophage lambda Q protein.
    Cold Spring Harb Symp Quant Biol. 1998;63:319-25 PMID: 10384296
  62. Structure and mechanism in transcriptional antitermination by the bacteriophage lambda N protein.
    Cold Spring Harb Symp Quant Biol. 1998;63:327-36 PMID: 10384297
  63. Promoter-associated pausing in promoter architecture and postinitiation transcriptional regulation.
    Cold Spring Harb Symp Quant Biol. 1998;63:347-56 PMID: 10384299
  64. The SAGA of Spt proteins and transcriptional analysis in yeast: past, present, and future.
    Cold Spring Harb Symp Quant Biol. 1998;63:553-61 PMID: 10384320
  65. A protein phosphatase functions to recycle RNA polymerase II.
    Genes Dev. 1999 Jun 15;13(12):1540-52 PMID: 10385623
  66. A novel RNA polymerase II-containing complex potentiates Tat-enhanced HIV-1 transcription.
    EMBO J. 1999 Jul 1;18(13):3688-701 PMID: 10393184
  67. Mechanism and regulation of transcriptional elongation by RNA polymerase II.
    Curr Opin Cell Biol. 1999 Jun;11(3):342-6 PMID: 10395562
  68. Coupling RNA polymerase II transcription with pre-mRNA processing.
    Curr Opin Cell Biol. 1999 Jun;11(3):347-51 PMID: 10395561
  69. The chromatin-specific transcription elongation factor FACT comprises human SPT16 and SSRP1 proteins.
    Nature. 1999 Jul 15;400(6741):284-8 PMID: 10421373
  70. Transcription elongation factor hSPT5 stimulates mRNA capping.
    Genes Dev. 1999 Jul 15;13(14):1774-9 PMID: 10421630
  71. The helix-loop-helix proteins dAP-4 and daughterless bind both in vitro and in vivo to SEBP3 sites required for transcriptional activation of the Drosophila gene Sgs-4.
    J Mol Biol. 1999 Aug 6;291(1):71-82 PMID: 10438607
  72. HIV-1 Tat: coping with negative elongation factors.
    Curr Opin Immunol. 1999 Aug;11(4):460-5 PMID: 10448148
  73. Tat-SF1 protein associates with RAP30 and human SPT5 proteins.
    Mol Cell Biol. 1999 Sep;19(9):5960-8 PMID: 10454543
  74. SMRTER, a Drosophila nuclear receptor coregulator, reveals that EcR-mediated repression is critical for development.
    Mol Cell. 1999 Aug;4(2):175-86 PMID: 10488333
  75. The NusA and NusG proteins of Escherichia coli increase the in vitro readthrough frequency of a transcriptional attenuator preceding the gene for the beta subunit of RNA polymerase.
    J Biol Chem. 1992 Jan 25;267(3):1449-54 PMID: 1370474
  76. Requirement for E. coli NusG protein in factor-dependent transcription termination.
    Cell. 1992 Mar 6;68(5):989-94 PMID: 1547498
  77. NusG, a new Escherichia coli elongation factor involved in transcriptional antitermination by the N protein of phage lambda.
    J Biol Chem. 1992 Mar 25;267(9):6012-9 PMID: 1532577
  78. Stability of Drosophila RNA polymerase II elongation complexes in vitro.
    Mol Cell Biol. 1992 May;12(5):2067-77 PMID: 1373806
  79. Control of formation of two distinct classes of RNA polymerase II elongation complexes.
    Mol Cell Biol. 1992 May;12(5):2078-90 PMID: 1569941
  80. A transcriptional switch between the Pig-1 and Sgs-4 genes of Drosophila melanogaster.
    Mol Cell Biol. 1993 Jan;13(1):184-95 PMID: 8417325
  81. Cytostellin: a novel, highly conserved protein that undergoes continuous redistribution during the cell cycle.
    J Cell Sci. 1992 Oct;103 ( Pt 2):381-8 PMID: 1478941
  82. A multisubunit complex associated with the RNA polymerase II CTD and TATA-binding protein in yeast.
    Cell. 1993 Jul 2;73(7):1361-75 PMID: 8324825
  83. The Drosophila fork head factor directly controls larval salivary gland-specific expression of the glue protein gene Sgs3.
    Nucleic Acids Res. 1996 Jun 15;24(12):2387-94 PMID: 8710511
Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
0890-9369
DOI
10.1101/gad.831900
Published
2000-10-15
Pages
2623-34
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC316994
Subset
IM
Grants
NIGMS NIH HHS · R01 GM032967 · United States
NIGMS NIH HHS · R37 GM032967 · United States
NIGMS NIH HHS · GM32967 · United States
NIGMS NIH HHS · GMOD5582 · United States
Databases
GENBANK
AF222864
External Links
PubMed source DOI: 10.1101/gad.831900 PMC Full Text View journal details
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