Nucleic Acids Research, 1994, Vol. 22, No. 16 3331-3338
© 1994
MOLECULAR BIOLOGY |
Minor groove contacts are essential for an interaction of the human cytomegalovirus IE2 protein with its DNA target
Institut für Klinische und Molekulare Virologie der Universität Erlangen-Nürnberg SchloGgarten 4, 91054 Erlangen, Germany
*To whom correspondence should be addressed
Received June 13, 1994. Revised July 26, 1994. Accepted July 26, 1994.
The 86 kDa immediate early-2 protein (IE2, IE86) of human cytomegalovirus (HCMV) is a multifunctional polypeptide that can regulate gene expression both ular, it represses its own mRNA synthesis by binding directly to a sequence element, termed cis repression signal (CRS), that is located between the TATA box and the transcriptional start site of the major IE enhancer/promoter of HCMV. Here, we provide evidence that IE86, unlike most sequence-specific DNA-binding proteins, interacts primarily within the minor groove of the DNA helix. This was shown by hydroxyl radical and methylation interference assays. In addition, binding studies with inosine-substituted oligonucleotides which have an altered major groove morphology without changing the surface of the minor groove, confirmed the results obtained in interference analyses. This establishes IE86 as a member of a small group of DNA binding proteins that interact with A-T rich sequences within the minor groove and which also includes the TATA-box binding protein TBP. Remarkably, IE86 and TBP are able to bind simultaneously in an immediate vicinity at the major IE enhancer/promoter of HCMV. As minor groove binding proteins are known to bend DNA heavily this could contribute to the observed negative regulation of transcription by IE86.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  R. L. Sanders, C. L. Clark, C. S. Morello, and D. H. Spector Development of Cell Lines That Provide Tightly Controlled Temporal Translation of the Human Cytomegalovirus IE2 Proteins for Complementation and Functional Analyses of Growth-Impaired and Nonviable IE2 Mutant Viruses J. Virol., July 15, 2008; 82(14): 7059 - 7077. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Sourvinos, N. Tavalai, A. Berndt, D. A. Spandidos, and T. Stamminger Recruitment of Human Cytomegalovirus Immediate-Early 2 Protein onto Parental Viral Genomes in Association with ND10 in Live-Infected Cells J. Virol., September 15, 2007; 81(18): 10123 - 10136. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. A. White, C. J. Del Rosario, R. L. Sanders, and D. H. Spector The IE2 60-Kilodalton and 40-Kilodalton Proteins Are Dispensable for Human Cytomegalovirus Replication but Are Required for Efficient Delayed Early and Late Gene Expression and Production of Infectious Virus J. Virol., March 15, 2007; 81(6): 2573 - 2583. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Reeves, J. Murphy, R. Greaves, J. Fairley, A. Brehm, and J. Sinclair Autorepression of the Human Cytomegalovirus Major Immediate-Early Promoter/Enhancer at Late Times of Infection Is Mediated by the Recruitment of Chromatin Remodeling Enzymes by IE86. J. Virol., October 1, 2006; 80(20): 9998 - 10009. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Asmar, L. Wiebusch, M. Truss, and C. Hagemeier The Putative Zinc Finger of the Human Cytomegalovirus IE2 86-Kilodalton Protein Is Dispensable for DNA Binding and Autorepression, Thereby Demarcating a Concise Core Domain in the C Terminus of the Protein J. Virol., November 1, 2004; 78(21): 11853 - 11864. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. A. White, C. L. Clark, V. Sanchez, and D. H. Spector Small Internal Deletions in the Human Cytomegalovirus IE2 Gene Result in Nonviable Recombinant Viruses with Differential Defects in Viral Gene Expression J. Virol., February 15, 2004; 78(4): 1817 - 1830. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Stamminger, M. Gstaiger, K. Weinzierl, K. Lorz, M. Winkler, and W. Schaffner Open Reading Frame UL26 of Human Cytomegalovirus Encodes a Novel Tegument Protein That Contains a Strong Transcriptional Activation Domain J. Virol., April 16, 2002; 76(10): 4836 - 4847. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Sanchez, C. L. Clark, J. Y. Yen, R. Dwarakanath, and D. H. Spector Viable Human Cytomegalovirus Recombinant Virus with an Internal Deletion of the IE2 86 Gene Affects Late Stages of Viral Replication J. Virol., February 22, 2002; 76(6): 2973 - 2989. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. A. Bryant, P. Mixon, M. Davidson, A. J. Bannister, T. Kouzarides, and J. H. Sinclair The Human Cytomegalovirus 86-Kilodalton Major Immediate-Early Protein Interacts Physically and Functionally with Histone Acetyltransferase P/CAF J. Virol., August 15, 2000; 74(16): 7230 - 7237. [Abstract] [Full Text]
|
|
|
|
|
|
A. Angulo, D. Kerry, H. Huang, E.-M. Borst, A. Razinsky, J. Wu, U. Hobom, M. Messerle, and P. Ghazal Identification of a Boundary Domain Adjacent to the Potent Human Cytomegalovirus Enhancer That Represses Transcription of the Divergent UL127 Promoter J. Virol., March 15, 2000; 74(6): 2826 - 2839. [Abstract] [Full Text]
|
|
|
|
|
|
S. Ghosh, A. Jain, B. Mukherjee, S. Habib, and S. E. Hasnain The Host Factor Polyhedrin Promoter Binding Protein (PPBP) Is Involved in Transcription from the Baculovirus Polyhedrin Gene Promoter J. Virol., September 1, 1998; 72(9): 7484 - 7493. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W. A. Bresnahan, T. Albrecht, and E. A. Thompson The Cyclin E Promoter Is Activated by Human Cytomegalovirus 86-kDa Immediate Early Protein J. Biol. Chem., August 21, 1998; 273(34): 22075 - 22082. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. J. Biegalke Characterization of the Transcriptional Repressive Element of the Human Cytomegalovirus Immediate-Early US3 Gene J. Virol., July 1, 1998; 72(7): 5457 - 5463. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. M. Rodems, C. L. Clark, and D. H. Spector Separate DNA Elements Containing ATF/CREB and IE86 Binding Sites Differentially Regulate the Human Cytomegalovirus UL112-113 Promoter at Early and Late Times in the Infection J. Virol., April 1, 1998; 72(4): 2697 - 2707. [Abstract] [Full Text] [PDF]
|
|