Structural and functional analyses of PAS domain interactions of the clock proteins Drosophila PERIOD and mouse PERIOD2

Sven Hennig, Holger M Strauss, Katja Vanselow, Ozkan Yildiz, Sabrina Schulze, Julia Arens, Achim Kramer, Eva Wolf

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

PERIOD proteins are central components of the Drosophila and mammalian circadian clocks. The crystal structure of a Drosophila PERIOD (dPER) fragment comprising two PER-ARNT-SIM (PAS) domains (PAS-A and PAS-B) and two additional C-terminal alpha-helices (alphaE and alphaF) has revealed a homodimer mediated by intermolecular interactions of PAS-A with tryptophane 482 in PAS-B and helix alphaF. Here we present the crystal structure of a monomeric PAS domain fragment of dPER lacking the alphaF helix. Moreover, we have solved the crystal structure of a PAS domain fragment of the mouse PERIOD homologue mPER2. The mPER2 structure shows a different dimer interface than dPER, which is stabilized by interactions of the PAS-B beta-sheet surface including tryptophane 419 (equivalent to Trp482dPER). We have validated and quantitatively analysed the homodimer interactions of dPER and mPER2 by site-directed mutagenesis using analytical gel filtration, analytical ultracentrifugation, and co-immunoprecipitation experiments. Furthermore we show, by yeast-two-hybrid experiments, that the PAS-B beta-sheet surface of dPER mediates interactions with TIMELESS (dTIM). Our study reveals quantitative and qualitative differences between the homodimeric PAS domain interactions of dPER and its mammalian homologue mPER2. In addition, we identify the PAS-B beta-sheet surface as a versatile interaction site mediating mPER2 homodimerization in the mammalian system and dPER-dTIM heterodimer formation in the Drosophila system.

Original languageEnglish
Pages (from-to)e94
JournalPloS Biology
Volume7
Issue number4
DOIs
Publication statusPublished - 28 Apr 2009

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Drosophila
Crystal structure
mice
Mutagenesis
proteins
crystal structure
Dimers
Yeast
Clocks
Gels
Experiments
Drosophila And protein
Drosophila Clk protein
Mouse Clock protein
Circadian Clocks
Proteins
ultracentrifugation
Ultracentrifugation
site-directed mutagenesis
Site-Directed Mutagenesis

Keywords

  • Animals
  • Biological Clocks
  • Cell Cycle Proteins
  • Circadian Rhythm
  • Drosophila
  • Drosophila Proteins
  • Mice
  • Nuclear Proteins
  • Period Circadian Proteins
  • Protein Multimerization
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • Sequence Alignment
  • Transcription Factors
  • Tryptophan
  • Journal Article
  • Research Support, Non-U.S. Gov't

Cite this

Hennig, Sven ; Strauss, Holger M ; Vanselow, Katja ; Yildiz, Ozkan ; Schulze, Sabrina ; Arens, Julia ; Kramer, Achim ; Wolf, Eva. / Structural and functional analyses of PAS domain interactions of the clock proteins Drosophila PERIOD and mouse PERIOD2. In: PloS Biology. 2009 ; Vol. 7, No. 4. pp. e94.
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Structural and functional analyses of PAS domain interactions of the clock proteins Drosophila PERIOD and mouse PERIOD2. / Hennig, Sven; Strauss, Holger M; Vanselow, Katja; Yildiz, Ozkan; Schulze, Sabrina; Arens, Julia; Kramer, Achim; Wolf, Eva.

In: PloS Biology, Vol. 7, No. 4, 28.04.2009, p. e94.

Research output: Contribution to JournalArticleAcademicpeer-review

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AU - Hennig, Sven

AU - Strauss, Holger M

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AB - PERIOD proteins are central components of the Drosophila and mammalian circadian clocks. The crystal structure of a Drosophila PERIOD (dPER) fragment comprising two PER-ARNT-SIM (PAS) domains (PAS-A and PAS-B) and two additional C-terminal alpha-helices (alphaE and alphaF) has revealed a homodimer mediated by intermolecular interactions of PAS-A with tryptophane 482 in PAS-B and helix alphaF. Here we present the crystal structure of a monomeric PAS domain fragment of dPER lacking the alphaF helix. Moreover, we have solved the crystal structure of a PAS domain fragment of the mouse PERIOD homologue mPER2. The mPER2 structure shows a different dimer interface than dPER, which is stabilized by interactions of the PAS-B beta-sheet surface including tryptophane 419 (equivalent to Trp482dPER). We have validated and quantitatively analysed the homodimer interactions of dPER and mPER2 by site-directed mutagenesis using analytical gel filtration, analytical ultracentrifugation, and co-immunoprecipitation experiments. Furthermore we show, by yeast-two-hybrid experiments, that the PAS-B beta-sheet surface of dPER mediates interactions with TIMELESS (dTIM). Our study reveals quantitative and qualitative differences between the homodimeric PAS domain interactions of dPER and its mammalian homologue mPER2. In addition, we identify the PAS-B beta-sheet surface as a versatile interaction site mediating mPER2 homodimerization in the mammalian system and dPER-dTIM heterodimer formation in the Drosophila system.

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KW - Biological Clocks

KW - Cell Cycle Proteins

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KW - Drosophila

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KW - Mice

KW - Nuclear Proteins

KW - Period Circadian Proteins

KW - Protein Multimerization

KW - Protein Structure, Secondary

KW - Protein Structure, Tertiary

KW - Sequence Alignment

KW - Transcription Factors

KW - Tryptophan

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

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JO - PloS Biology

JF - PloS Biology

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