Two Protocols to Study the Interactions of Thyroid Hormone Receptors with Other Proteins and Chromatin

Henrik Holbech , Peter Matthiessen , Martin Hansen , Gerrit Schüürmann , Dries Knapen , Marieke Reuver , Laurent Sachs , Werner Kloas , Klara Hilscherova , Marc Leonard , Jürgen Arning , Volker Strauss , Taisen Iguchi , Lisa Baumann , Albane Le Maire , Natacha Bouhours-Nouet , Jessica Soamalala , Delphine Mirebeau-Prunier , Matteo Paloni , Laura Guee , Delphine Heron , Cyril Mignot , Frédéric Illouz , Florence Joubert , Claire Briet , Patrice Rodien , William Bourguet , Hui Sun , Haiying Wu , Rongrong Xie , Fengyun Wang , Ting Chen , Xiuli Chen , Xiaoyan Wang , Linqi Chen , Wenyue Guan , Romain Guyot , Sabine Richard , Frederic Flamant (1)

1 IGFL - Institut de Génomique Fonctionnelle de Lyon

Understanding the transcriptional function of thyroid hormone receptors implies a precise analysis of their interactions with chromatin and other protein components of the cells. We present here two protocols that are routinely used in our laboratory. The first co-immunoprecipitation procedure allows addressing the capacity of proteins to form stable multiprotein complexes with TRs in cells. The chromatin affinity purification enables us to define the sites occupied by TRs on chromatin. In this case the lack of high quality antibodies is circumvented by introducing an N-terminal tag in TR, with unspecific affinity for immunoglobulins.

Mots clés: AOP; IATA; OECD; adverse outcome pathway; biomarkers; cross-species extrapolation; endocrine disruption; integrated approach to testing and assessment; test guideline; thyroid hormone disruption; genetic disease; nuclear receptor; thyroid hormone; Chromatin affinity purification; Co-immunoprecipitation; Thyroid hormone receptors; brain–blood barrier; neurodevelopment; neurodevelopmental disorder; placenta; thyroid hormones; transporters

Domaines: Biologie du développement

Henrik Holbech, Peter Matthiessen, Martin Hansen, Gerrit Schüürmann, Dries Knapen, et al.. Two Protocols to Study the Interactions of Thyroid Hormone Receptors with Other Proteins and Chromatin. Frontiers in Endocrinology, 2018, 1801 (3), pp.9-16. ⟨10.1007/978-1-4939-7902-8_2⟩. ⟨hal-03278376⟩