Krüppel-like factor 11 regulates the expression of metabolic genes via an evolutionarily conserved protein interaction domain functionally disrupted in maturity onset diabetes of the young.

Publication Type:

Journal Article

Source:

The Journal of biological chemistry, Volume 288, Issue 24, p.17745-58 (2013)

Keywords:

Amino Acid Sequencedigestive disease, digestive deseases Animalsdigestive disease, digestive deseases Cell Cycle Proteinsdigestive disease, digestive deseases CHO Cellsdigestive disease, digestive deseases Conserved Sequencedigestive disease, digestive deseases Cricetinaedigestive disease, digestive deseases Diabetes Mellitus, Type 2digestive disease, digestive deseases Evolution, Moleculardigestive disease, digestive deseases Gene Expression Regulationdigestive disease, digestive deseases Gene Regulatory Networksdigestive disease, digestive deseases Glucosedigestive disease, digestive deseases GTP-Binding Protein beta Subunitsdigestive disease, digestive deseases Humansdigestive disease, digestive deseases Insulindigestive disease, digestive deseases Molecular Sequence Datadigestive disease, digestive deseases Mutation, Missensedigestive disease, digestive deseases Promoter Regions, Geneticdigestive disease, digestive deseases Protein Bindingdigestive disease, digestive deseases Protein Interaction Domains and Motifsdigestive disease, digestive deseases Ratsdigestive disease, digestive deseases Repressor Proteinsdigestive disease, digestive deseases Signal Transductiondigestive disease, digestive deseases Transcription, Geneticdigestive disease, digestive deseases Two-Hybrid System Techniques

Abstract:

The function of Krüppel-like factor 11 (KLF11) in the regulation of metabolic pathways is conserved from flies to human. Alterations in KLF11 function result in maturity onset diabetes of the young 7 (MODY7) and neonatal diabetes; however, the mechanisms underlying the role of this protein in metabolic disorders remain unclear. Here, we investigated how the A347S genetic variant, present in MODY7 patients, modulates KLF11 transcriptional activity. A347S affects a previously identified transcriptional regulatory domain 3 (TRD3) for which co-regulators remain unknown. Structure-oriented sequence analyses described here predicted that the KLF11 TRD3 represents an evolutionarily conserved protein domain. Combined yeast two-hybrid and protein array experiments demonstrated that the TRD3 binds WD40, WWI, WWII, and SH3 domain-containing proteins. Using one of these proteins as a model, guanine nucleotide-binding protein β2 (Gβ2), we investigated the functional consequences of KLF11 coupling to a TRD3 binding partner. Combined immunoprecipitation and biomolecular fluorescence complementation assays confirmed that activation of three different metabolic G protein-coupled receptors (β-adrenergic, secretin, and cholecystokinin) induces translocation of Gβ2 to the nucleus where it directly binds KLF11 in a manner that is disrupted by the MODY7 A347S variant. Using genome-wide expression profiles, we identified metabolic gene networks impacted upon TRD3 disruption. Furthermore, A347S disrupted KLF11-mediated increases in basal insulin levels and promoter activity and blunted glucose-stimulated insulin secretion. Thus, this study characterizes a novel protein/protein interaction domain disrupted in a KLF gene variant that associates to MODY7, contributing to our understanding of gene regulation events in complex metabolic diseases.