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The Human KROX-26/ZNF22 Gene is Expressed at Sites of Tooth Formation and Maps to the Locus for Permanent Tooth Agenesis (He-Zhao Deficiency)

¹ Canadian Institutes for Health Research (CIHR) Group in Matrix Dynamics, University of Toronto, Faculty of Dentistry, Fitzgerald Building, Room 239, 150 College Street, Toronto, ON M5S 3E2, Canada; and
² Tokyo Medical and Dental University, Department of Hard Tissue Engineering (Periodontology), Bunkyo-ku, Tokyo, Japan;

View larger version (127K): [in a new window]   Figure 1. KROX-26 genomic DNA sequence. The NarI/Bst1107I genomic DNA fragment from BAC clone RP11-285G1 contains the entire KROX-26 open reading frame (underlined) as well as 2.4 kb of upstream sequence and 1.3 kb of downstream sequence. Several TATA and CCAAT box consensus sites in the upstream region are shown in bold, and the conserved polyadenylation site in the 3'-untranslated region is underlined. The sequence differences between KROX-26 and the HKR-T1 cDNA sequence are indicated in bold and italicized lower case in positions 2517, 2518, and 3074.

View larger version (35K): [in a new window]   Figure 2. Sequence alignment between mouse and human Krox-26 protein sequences. The zinc finger region in both proteins is indicated in bold. Conserved amino acids are indicated by asterisks, highly similar amino acid substitutions by colons, and conserved amino acid substitutions by periods. The highest degree of sequence conservation is found in the zinc finger region.

View larger version (125K): [in a new window]   Figure 3. KROX-26 mRNA expression in embryonic (A) and adult (B) human tissues. Tissue sections from 12-week-old human embryos were hybridized with a digoxigenin-labeled KROX-26 antisense (a, c, e, g, i, k) or sense (b, d, f, h, j, l) probe and counterstained with methyl green. Positive signals in blue or brown (arrows), indicating KROX-26 expression, are seen in the epithelial component of the developing tooth (a, c, e), tongue epithelium (g), tongue striated muscle (i), and osteoblasts of craniofacial bone (k). Bars indicate 250 µm (a-f) or 100 µm (g-l). Abbreviations: B, bone; E, epithelium; M, mesenchyme. A multiple-tissue Northern Blot on human adult tissues (B) indicates the existence of a single 2.4-kb KROX-26 mRNA transcript in multiple tissues at various levels, relative to ß-actin mRNA expression.

View larger version (53K): [in a new window]   Figure 4. Chromosomal localization of the KROX-26 gene by FISH. Hybridization of a biotinylated KROX-26 DNA probe to human metaphase chromosome spreads and detection with FITC revealed signals on both alleles of chromosome 10 in position 10q11.21 (arrows).