title: Identification of human ccn2 (connective tissue growth factor) promoter polymorphisms.
abstract: BACKGROUND
  Connective tissue growth factor (CCN2; CTGF) is a newly identified growth factor, which is involved in the regulation of wound repair and fibrosis. Because there is variation among individuals with respect to tissue response to injury, genetic factors might be involved in the final outcome of tissue repair or scarring. For example, polymorphisms in the promoter region of genes, such as those encoding transforming growth factor beta1 (TGF-beta1), interleukin 10 (IL-10), and tumour necrosis factor alpha (TNF-alpha), influence transcriptional responses and are thought to contribute to the dysregulation of these genes in pathological conditions.
  
  
  OBJECTIVE
  To investigate whether the promoter region of the ccn2 (ctgf) gene contains polymorphic sequences that might account for differential expression.
  
  
  METHODS
  Seventy seven human DNA samples were sequenced-45 were from healthy controls and 32 were from patients with ischaemic heart disease (IHD)-using M13 tailed sequence specific ccn2 (ctgf) primers for amplification of a 600 bp fragment upstream of the transcription start site. Amplicons were bidirectionally sequenced with a dye primer M13 forward and reverse sequencing kit.
  
  
  RESULTS
  A C to G substitution was identified at position -132 in one of the patients with IHD. Moreover, in five of the 32 patients with IHD and in six of the 45 healthy controls, a G to C polymorphism was found at position -447. These substitutions at -132 and -447 are thought to lie within predicted binding domains for the transcription factors Pbx-1 and MZF1, respectively. In addition, insertions at position -43 (G), -47 (C), -71 (G) and a C to T substitution at position -198 were found in all DNA samples compared with the published ccn2 (ctgf) promoter sequence. These corrections do not involve sequences predicted to function as transcription factor binding sites.
  
  
  CONCLUSIONS
  Sequence analysis of the ccn2 (ctgf) promoter of 77 human DNA samples has revealed corrections and polymorphic sites. The latter lie within putative regulatory elements.
authors.affiliation: Department of Pathology, H04.312, University Medical Centre Utrecht, Heidelberglaan 100, 3584 CX Utrecht, The Netherlands.
authors.forename: I E
authors.initials: IE
authors.lastname: Blom
authors.forename: A J
authors.initials: AJ
authors.lastname: van Dijk
authors.forename: R A
authors.initials: RA
authors.lastname: de Weger
authors.forename: M G
authors.initials: MG
authors.lastname: Tilanus
authors.forename: R
authors.initials: R
authors.lastname: Goldschmeding
mesh_terms: D000483:Alleles; D019943:Amino Acid Substitution / Q000235:genetics; D001665:Binding Sites; D016022:Case-Control Studies; D055513:Connective Tissue Growth Factor; D005838:Genotype; D006133:Growth Substances / Q000235:genetics*; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics*; D036341:Intercellular Signaling Peptides and Proteins; D017202:Myocardial Ischemia / Q000235:genetics; D011110:Polymorphism, Genetic; D011401:Promoter Regions, Genetic; D017422:Sequence Analysis, DNA; D012984:Software; D014157:Transcription Factors / Q000235:genetics
chemical_list: C526317:CCN2 protein, human; D006133:Growth Substances; D017874:Immediate-Early Proteins; D036341:Intercellular Signaling Peptides and Proteins; D014157:Transcription Factors; D055513:Connective Tissue Growth Factor