Positional Cloning of Zinc Finger Domain Transcription Factor Zfp69, a Candidate Gene for Obesity-Associated Diabetes Contributed by Mouse Locus Nidd/SJL

Polygenic type 2 diabetes in mouse models is associated with obesity and results from a combination of adipogenic and diabetogenic alleles. Here we report the identification of a candidate gene for the diabetogenic effect of a QTL (Nidd/SJL, Nidd1) contributed by the SJL, NON, and NZB strains in outcross populations with New Zealand Obese (NZO) mice. A critical interval of distal chromosome 4 (2.1 Mbp) conferring the diabetic phenotype was identified by interval-specific congenic introgression of SJL into diabetes-resistant C57BL/6J, and subsequent reporter cross with NZO. Analysis of the 10 genes in the critical interval by sequencing, qRT–PCR, and RACE–PCR revealed a striking allelic variance of Zfp69 encoding zinc finger domain transcription factor 69. In NZO and C57BL/6J, a retrotransposon (IAPLTR1a) in intron 3 disrupted the gene by formation of a truncated mRNA that lacked the coding sequence for the KRAB (Krüppel-associated box) and Znf-C2H2 domains of Zfp69, whereas the diabetogenic SJL, NON, and NZB alleles generated a normal mRNA. When combined with the B6.V-Lepob background, the diabetogenic Zfp69SJL allele produced hyperglycaemia, reduced gonadal fat, and increased plasma and liver triglycerides. mRNA levels of the human orthologue of Zfp69, ZNF642, were significantly increased in adipose tissue from patients with type 2 diabetes. We conclude that Zfp69 is the most likely candidate for the diabetogenic effect of Nidd/SJL, and that retrotransposon IAPLTR1a contributes substantially to the genetic heterogeneity of mouse strains. Expression of the transcription factor in adipose tissue may play a role in the pathogenesis of type 2 diabetes.

[1]  M. McCarthy,et al.  Replication of Genome-Wide Association Signals in UK Samples Reveals Risk Loci for Type 2 Diabetes , 2007, Science.

[2]  E. Leiter,et al.  Deconstructing and reconstructing obesity-induced diabetes (diabesity) in mice. , 2002, Diabetes.

[3]  S. Henikoff,et al.  Predicting deleterious amino acid substitutions. , 2001, Genome research.

[4]  D. Speicher,et al.  The structurally disordered KRAB repression domain is incorporated into a protease resistant core upon binding to KAP-1-RBCC domain. , 2007, Journal of molecular biology.

[5]  M. Satake,et al.  SMAP2, a novel ARF GTPase-activating protein, interacts with clathrin and clathrin assembly protein and functions on the AP-1-positive early endosome/trans-Golgi network. , 2006, Molecular biology of the cell.

[6]  J. Berthelsen,et al.  Purification and Characterization of UEF3, A Novel Factor Involved in the Regulation of the Urokinase and Other AP-1 Controlled Promoters (*) , 1996, The Journal of Biological Chemistry.

[7]  E. Leiter,et al.  NIDDM Genes in Mice: Deleterious Synergism by Both Parental Genomes Contributes to Diabetogenic Thresholds , 1998, Diabetes.

[8]  J. Martial,et al.  The evolutionarily conserved Krüppel-associated box domain defines a subfamily of eukaryotic multifingered proteins. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[9]  Kang Z. Liu,et al.  Intronic polyadenylation signal sequences and alternate splicing generate human soluble Fltl variants and regulate the abundance of soluble Flt1 in the placenta , 2007, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[10]  Philipp W. Raess,et al.  Positional cloning of Sorcs1, a type 2 diabetes quantitative trait locus , 2006, Nature Genetics.

[11]  E. Bouhassira,et al.  From bloodjournal.hematologylibrary.org at PENN STATE UNIVERSITY on February 23, 2013. For personal use only. , 2007 .

[12]  T. Hudson,et al.  A genome-wide association study identifies novel risk loci for type 2 diabetes , 2007, Nature.

[13]  B. Kahn.  Type 2 Diabetes: When Insulin Secretion Fails to Compensate for Insulin Resistance , 1998, Cell.

[14]  Marcia M. Nizzari,et al.  Genome-Wide Association Analysis Identifies Loci for Type 2 Diabetes and Triglyceride Levels , 2007, Science.

[15]  M. Yanagisawa,et al.  Reduced Expression of the Endothelin Receptor Type B Gene in Piebald Mice Caused by Insertion of a Retroposon-like Element in Intron 1* , 2006, Journal of Biological Chemistry.

[16]  M. Heiman,et al.  Evaluation of a quantitative magnetic resonance method for mouse whole body composition analysis. , 2004, Obesity research.

[17]  L. Plum,et al.  Type 2 diabetes-like hyperglycemia in a backcross model of NZO and SJL mice: characterization of a susceptibility locus on chromosome 4 and its relation with obesity. , 2000, Diabetes.

[18]  J. Gulcher,et al.  A variant in CDKAL1 influences insulin response and risk of type 2 diabetes , 2007, Nature Genetics.

[19]  Raul Urrutia,et al.  KRAB-containing zinc-finger repressor proteins , 2003, Genome Biology.

[20]  W. Chung,et al.  Positional Cloning of “Lisch-like”, a Candidate Modifier of Susceptibility to Type 2 Diabetes in Mice , 2008, PLoS genetics.

[21]  Rob C Hoeben,et al.  Tbc1d1 mutation in lean mouse strain confers leanness and protects from diet-induced obesity , 2008, Nature Genetics.

[22]  G. Abecasis,et al.  A Genome-Wide Association Study of Type 2 Diabetes in Finns Detects Multiple Susceptibility Variants , 2007, Science.

[23]  J. Schuchhardt,et al.  A meta-analysis of QTL for diabetes-related traits in rodents. , 2008, Physiological genomics.

[24]  T. Aigner,et al.  Collagens--structure, function, and biosynthesis. , 2003, Advanced drug delivery reviews.

[25]  D. Coleman,et al.  Laboratory animals exhibiting obesity and diabetes syndromes. , 1977, Metabolism: clinical and experimental.

[26]  J. Lawler,et al.  Skeletal abnormalities in mice lacking extracellular matrix proteins, thrombospondin-1, thrombospondin-3, thrombospondin-5, and type IX collagen. , 2008, The American journal of pathology.

[27]  D. Altshuler,et al.  The inherited basis of diabetes mellitus: implications for the genetic analysis of complex traits. , 2003, Annual review of genomics and human genetics.

[28]  A. Schürmann,et al.  Characterisation of the mouse diabetes susceptibility locus Nidd/SJL: islet cell destruction, interaction with the obesity QTL Nob1, and effect of dietary fat , 2002, Diabetologia.

[29]  E. Leiter,et al.  Obesity/Diabetes in Mice with Mutations in Leptin or Leptin Receptor Genes , 2007 .

[30]  Thomas D. Schmittgen,et al.  Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. , 2001, Methods.

[31]  A. Schürmann,et al.  ADP-ribosylation Factor-like GTPase ARFRP1 Is Required for Trans-Golgi to Plasma Membrane Trafficking of E-cadherin* , 2008, Journal of Biological Chemistry.

[32]  C. Bogardus,et al.  Evidence for Linkage Between a Region on Chromosome 1p and the Acute Insulin Response in Pima Indians , 1995, Diabetes.

[33]  Ying Zhang,et al.  Genome-Wide Assessments Reveal Extremely High Levels of Polymorphism of Two Active Families of Mouse Endogenous Retroviral Elements , 2008, PLoS genetics.