NKX2-5: An Update on this Hypermutable Homeodomain Protein and its Role in Human Congenital Heart Disease (CHD) Stella Marie Reamon-Buettner, Juergen T Borlak To cite this version: Stella Marie Reamon-Buettner, Juergen T Borlak. NKX2-5: An Update on this Hypermutable Home- odomain Protein and its Role in Human Congenital Heart Disease (CHD). Human Mutation, Wiley, 2010, 31 (11), pp.1185. 10.1002/humu.21345. hal-00585168 HAL Id: hal-00585168 https://hal.archives-ouvertes.fr/hal-00585168 Submitted on 12 Apr 2011 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Human Mutation NKX2-5: An Update on this Hypermutable Homeodomain Protein and its Role in Human Congenital Heart Disease (CHD) For Peer Review Journal: Human Mutation Manuscript ID: humu-2010-0256.R1 Wiley - Manuscript type: Review Date Submitted by the 15-Jul-2010 Author: Complete List of Authors: Reamon-Buettner, Stella Marie; Fraunhofer Institute of Toxicology and Experimental Medicine, Molecular Medicine and Medical Biotechnology Borlak, Juergen; Fraunhofer Institute of Toxicology and Experimental Medicine, Molecular Medicine and Medical Biotechnology heart development, congenital heart disease, cardiac Key Words: malformations, transcription factors, NKX2-5, mutations John Wiley & Sons, Inc. Page 1 of 59 Human Mutation 1 1 2 NKX2-5: An Update on this Hypermutable Homeodomain Protein 3 4 5 and its Role in Human Congenital Heart Disease (CHD) 6 7 8 9 Stella Marie Reamon-Buettner and Juergen Borlak* 10 11 12 13 14 15 Molecular Medicine and Medical Biotechnology, Fraunhofer Institute of Toxicology and 16 17 Experimental Medicine, Nikolai-Fuchs-Strasse 1, D-30625 Hannover, Germany. 18 19 20 For Peer Review 21 22 23 24 25 *Correspondence to Prof. Dr. Juergen Borlak, Molecular Medicine and Medical 26 27 Biotechnology, Fraunhofer Institute of Toxicology and Experimental Medicine, Nikolai- 28 29 Fuchs-Strasse 1, 30625 Hannover, Germany. 30 31 Tel: +49-511-5350-559; Fax +49-511-5350-573; E-mail: [email protected] 32 33 34 35 36 37 38 39 Contract grant sponsors: Ministry of Science and Culture, Lower Saxony, Germany; Contract 40 grant number: 25A.5-76251-99-3/00 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 John Wiley & Sons, Inc. Human Mutation Page 2 of 59 2 1 2 ABSTRACT 3 4 Congenital heart disease (CHD) is among the most prevalent and fatal of all birth defects. 5 6 Deciphering its causes, however, is complicated as many patients affected by CHD have no 7 8 family history of the disease. There is also widespread heterogeneity of cardiac malformations 9 Deleted : within 10 among affected individuals. Nonetheless, there have been tremendous efforts towards a better 11 12 understanding of the molecular and cellular events leading to CHD. Notably, certain cardiac- 13 14 specific transcription factors have been implicated in mammalian heart development and 15 Deleted : in CHD; foremost is 16 disruption of their activity has been demonstrated in CHD. The homeodomain transcription Deleted : t 17 18 factor NKX2-5 is an important member of this group . Indeed, more than 40 heterozygous 19 20 NKX2-5 germline mutations haveFor been observed Peer in individuals Review with CHD and these are 21 22 spread along the coding region, with many shown to impact protein function. Thus, NKX2-5 23 24 appears to be hypermutable (?) highly mutable , yet the overall detection frequency in sporadic 25 26 CHD is about 2% and NKX2-5 mutations are one-time detections with single-positives or 27 28 private to families. Furthermore, there is lack of genotype-phenotype correlation, in which the 29 30 same cardiac malformations have been exhibited in different NKX2-5 mutations or the same 31 32 NKX2-5 mutation associated with diverse malformations. Here, we summarize published 33 34 NKX2-5 germline mutations and explore different avenues in disease pathogenesis to support 35 36 the notion of a multifactorial cause of CHD where possibly several genes and associated 37 38 pathways are involved. 39 40 41 42 43 44 Keywords : heart development, congenital heart disease, cardiac malformations, transcription 45 46 factors, NKX2-5, mutations 47 Deleted : Page Break ¶ 48 Formatted: Don't adjust right 49 indent when grid is defined, Line 50 spacing: Double, Don't adjust 51 space between Latin and Asian text, Don't adjust space between 52 Asian text and numbers 53 54 55 56 57 58 59 60 John Wiley & Sons, Inc. Page 3 of 59 Human Mutation 1 1 2 3 Dr. Mark Paalman 4 5 The Managing Editor 6 Human Mutation 7 8 Dear Dr. Paalman: 9 10 Many thanks for the encouraging news on our manuscript " NKX2-5: An Update on this 11 12 Hypermutable Homeodomain Protein and its Role in Human Congenital Heart Disease (CHD)" 13 Human Mutation Paper No. humu-2010-0256. Attached is the revised manuscript, as well as the 14 response to the managing editor's comments. 15 16 17 We look forward to seeing our work published in Human Mutation. 18 For Peer Review 19 20 Sincerely yours, 21 22 JUERGEN BORLAK 23 24 25 26 27 RESPONSE TO MANAGING EDITOR'S COMMENTS 28 29 30 1) The mutation nomenclature appears to follow the format indicated in the 31 Author Instructions (see the website http://www.hgvs.org/mutnomen/ and also 32 the nomenclature checklist at http://www.hgvs.org/mutnomen/checklist.html). Things to watch for: 33 34 a-Mention the GenBank reference sequence and version number for the gene(s) 35 studied (i.e., include the decimal point following the accession number in the 36 sequence record) in the Materials and Methods and as a footnote to the 37 relevant tables and figure. 38 b-Clearly indicate in the Methods text and tables that the DNA mutation 39 numbering system you use is based on cDNA sequence. Explanatory text is 40 required in legends and text, for example: 41 “Nucleotide numbering reflects cDNA numbering with +1 corresponding to the A 42 of the ATG translation initiation codon in the reference sequence, according 43 to journal guidelines (www.hgvs.org/mutnomen). The initiation codon is codon 44 1.” 45 46 RESPONSE : 47 The footnote below is incorporated in Table 1, Supp.Tables S2 and S3. 48 49 50 "Nucleotide numbering reflects cDNA numbering with +1 corresponding to the A of the ATG translation initiation 51 codon in the reference sequence NM_0004387.3, according to journal guidelines ( www.hgvs.org/mutnomen ). The 52 initiation codon is codon 1. " 53 54 c-Authors are advised to check sequence variant descriptions using the 55 Mutalyzer program 56 (http://www.LOVD.nl/mutalyzer/) - using batch mode, all variants for a gene 57 can be analyzed at once. See the article Wildeman et al. Hum Mutat 29, 6-13, 58 2008 http://www3.interscience.wiley.com/cgi-bin/fulltext/116842325/PDFSTART 59 60 John Wiley & Sons, Inc. Human Mutation Page 4 of 59 2 1 2 3 4 5 RESPONSE : 6 The batch mode of Mutalyzer was used to check the list of NKX2-5 mutations. 7 8 d-All authors should verify that the mutations reported (especially novel 9 ones) have been or will be submitted to an existing locus-specific database 10 for the genes involved. Visit the HGVS-LSDB list to search for databases: 11 http://www.hgvs.org/dblist/glsdb.html 12 RESPONSE : 13 14 Review paper, and no novel mutations being reported. 15 16 2) On resubmission: a-Please double check the author names and affiliations carefully. These are 17 often a source of typographical errors. 18 For Peer Review 19 RESPONSE : 20 Authors' names and affiliations checked. 21 22 b-Title Page, Abstract (180-200 words max), Key Words, Main Text, References, 23 and Figure Legends should be combined into one file for the manuscript and 24 submitted as a *.doc file. 25 26 RESPONSE : 27 The submitted*doc. file (NKX2-5 humu10-0256_text.doc) contains the specified components; 28 Abstract is 200 words. 29 c-Figures for main article must be submitted as separate files with high 30 resolution (at least 200 dpi) as *.tif or *.eps format only. 31 --For color figures in print: submit two files for each color figure: one in 32 CMYK color space and one in RGB color space (with the true color you wish to 33 have published). If you cannot provide CMYK color e-files for the print version, please let me know. 34 35 RESPONSE : 36 Figure3 is 200 dpi, the rest are 300 dpi; both CMYK and RGB files for Figure 3 and Figure 4. 37 38 39 d-Tables must be submitted individually as separate *.doc files (with their 40 titles and legends included). Please use the MS Word table format if possible. 41 Excel (*.xls) files should not be submitted. 42 --Do not use custom paper sizes - only use Letter or A4. 43 RESPONSE : 44 45 Table 1. Functional Consequences of Identified NKX2-5 Mutations in CHD: Functional Assays 46 or Bioinformatic Analysis is contained in *doc.file NKX2-5 humu10-0256_Table 1.doc. 47 48 Table 2. Detection Frequency of NKX2-5 Germline Mutations in Congenital Heart Disease is 49 contained in *doc.file NKX2-5 humu10-0256_Table 2.doc.