title: 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.
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.
authors.affiliation: Laboratory of Epigenetics and Chromatin Dynamics, Epigenomics Translational Program, Mayo Clinic Center for Individualized Medicine, Division of Gastroenterology and Hepatology, and Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, Minnesota 55905, USA.
authors.forename: Gwen
authors.initials: G
authors.lastname: Lomberk
authors.forename: Adrienne
authors.initials: A
authors.lastname: Grzenda
authors.forename: Angela
authors.initials: A
authors.lastname: Mathison
authors.forename: Carlos
authors.initials: C
authors.lastname: Escande
authors.forename: Jin-San
authors.initials: JS
authors.lastname: Zhang
authors.forename: Ezequiel
authors.initials: E
authors.lastname: Calvo
authors.forename: Laurence J
authors.initials: LJ
authors.lastname: Miller
authors.forename: Juan
authors.initials: J
authors.lastname: Iovanna
authors.forename: Eduardo N
authors.initials: EN
authors.lastname: Chini
authors.forename: Martin E
authors.initials: ME
authors.lastname: Fernandez-Zapico
authors.forename: Raul
authors.initials: R
authors.lastname: Urrutia
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D051017:Apoptosis Regulatory Proteins; D016466:CHO Cells; D018797:Cell Cycle Proteins / Q000737:chemistry / Q000502:physiology*; D017124:Conserved Sequence; D006224:Cricetinae; D003924:Diabetes Mellitus, Type 2 / Q000235:genetics*; D019143:Evolution, Molecular; D044387:GTP-Binding Protein beta Subunits / Q000378:metabolism; D005786:Gene Expression Regulation; D053263:Gene Regulatory Networks; D005947:Glucose / Q000502:physiology; D006801:Humans; D007328:Insulin / Q000235:genetics / Q000378:metabolism; D000078790:Insulin Secretion; D008969:Molecular Sequence Data; D020125:Mutation, Missense; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D051381:Rats; D012097:Repressor Proteins / Q000737:chemistry / Q000502:physiology*; D015398:Signal Transduction; D014158:Transcription, Genetic; D020798:Two-Hybrid System Techniques
chemical_list: D051017:Apoptosis Regulatory Proteins; D018797:Cell Cycle Proteins; D044387:GTP-Binding Protein beta Subunits; D007328:Insulin; C115117:KLF11 protein, human; D012097:Repressor Proteins; D005947:Glucose
keywords: G Proteins; Gene Regulation; Gβ2; KLF11; Krüppel-like Factor (KLF); Metabolic Gene Networks; Protein-Protein Interactions; Transcription; WD40