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Solution NMR structure of MED25(391–543) comprising the activator-interacting domain (ACID) of human mediator subunit 25

  • Published:
Journal of Structural and Functional Genomics

Abstract

The solution NMR structure of protein MED25(391–543), comprising the activator interacting domain (ACID) of subunit 25 of the human mediator, is presented along with the measurement of polypeptide backbone heteronuclear 15N-{1H} NOEs to identify fast internal motional modes. This domain interacts with the acidic transactivation domains of Herpes simplex type 1 (HSV-1) protein VP16 and the Varicella-zoster virus (VZV) major transactivator protein IE62, which initiate transcription of viral genes. The structure is similar to the β-barrel domains of the human protein Ku and the SPOC domain of human protein SHARP, and provides a starting point to understand the structural biology of initiation of HSV-1 and VZV gene activation. Homology models built for the two ACID domains of the prostate tumor overexpressed (PTOV1) protein using the structure of MED25(391–543) as a template suggest that differential biological activities of the ACID domains in MED25 and PTOV1 arise from modulation of quite similar protein–protein interactions by variable residues grouped around highly conserved charged surface areas.

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Fig. 1
Fig. 2

Abbreviations

ACID:

Activator-interacting domain

CBP:

CREB-binding protein

HDAC:

Histone deacetylase complex

HSV-1:

Herpes simplex virus type 1

NESG:

Northeast Structural Genomics Consortium

MED25:

Subunit 25 of the human mediator complex

NOE:

Nuclear Overhauser effect

PDB:

Protein Data Bank

PTOV:

Prostate tumor overexpressed

RMSD:

Root mean square deviation

SHARP:

SMRT/HDAC1-associated repressor protein

SPOC:

Spen paralog and ortholog C-terminal domain

TAD:

Transactivation domain

VBD:

VP16-binding domain

VZV:

Varicella-zoster virus

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Acknowledgments

We thank R. Shastry, C. Ciccosanti, H. Janjua, and G.V.T. Swapna for contributions in sample preparation, and J. K. Everett and S. Bhattacharya for helpful discussions. This work was supported by the National Institutes of Health, grant numbers: U54 GM094597 (T.S. and G.T.M.) and R01 AI18449 (W.T.R.). Prof. T. Szyperski is a member of the New York Structural Biology Center. The Center is a STAR center supported by the New York State Office of Science, Technology, and Academic Research. 900 MHz spectrometer was purchased with funds from NIH, USA, the Keck Foundation, New York State, and the NYC Economic Development Corporation. Support was also obtained from the University at Buffalo’s Center for Computational Research.

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Authors and Affiliations

  1. Department of Chemistry, The State University of New York at Buffalo, Buffalo, NY, 14260, USA

    Alexander Eletsky & Thomas Szyperski

  2. Northeast Structural Genomics Consortium, Buffalo, NY, 14260, USA

    Alexander Eletsky & Thomas Szyperski

  3. Department of Microbiology and Immunology, The State University of New York at Buffalo, Buffalo, NY, 14214, USA

    William T. Ruyechan

  4. Northeast Structural Genomics Consortium, Buffalo, NY, 14214, USA

    William T. Ruyechan

  5. Center of Advanced Biotechnology and Medicine, Department of Molecular Biology and Biochemistry, Rutgers, The State University of New Jersey, Piscataway, NJ, 08854, USA

    Rong Xiao, Thomas B. Acton & Gaetano T. Montelione

  6. Department of Biochemistry, Robert Wood Johnson Medical School, University of Medicine and Dentistry of New Jersey, Piscataway, NJ, 08854, USA

    Rong Xiao, Thomas B. Acton & Gaetano T. Montelione

  7. Northeast Structural Genomics Consortium, Piscataway, NJ, 08854, USA

    Rong Xiao, Thomas B. Acton & Gaetano T. Montelione

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  1. Alexander Eletsky
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  2. William T. Ruyechan
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  3. Rong Xiao
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  5. Gaetano T. Montelione
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Correspondence to Thomas Szyperski.

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Eletsky, A., Ruyechan, W.T., Xiao, R. et al. Solution NMR structure of MED25(391–543) comprising the activator-interacting domain (ACID) of human mediator subunit 25. J Struct Funct Genomics 12, 159–166 (2011). https://doi.org/10.1007/s10969-011-9115-1

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