title: Deletion of a conserved noncoding sequence in Plzf intron leads to Plzf down-regulation in limb bud and polydactyly in the rat.
abstract: Lx mutation in SHR.Lx rat manifests in homozygotes as hindlimb preaxial polydactyly. It was previously mapped to a chromosome 8 segment containing the Plzf gene. Plzf (promyelocytic leukemia zinc finger protein) influences limb development as a direct repressor of posterior HoxD genes. However, the Plzf coding sequence is intact in the Lx mutants. Using linkage mapping in F2 hybrids, we downsized the segment containing Lx to 155 kb and sequenced conserved noncoding elements (CNEs) inside. A 2,964-bp deletion in Plzf intron 2, never detected in control animals, is the only candidate for Lx. The deletion removes the most deeply conserved CNE in the 155-kb segment, suggesting a regulatory influence on Plzf expression. Correspondingly, using in situ hybridization and quantitative real-time polymerase chain reaction, we found a decrease of Plzf expression in Lx/Lx limb buds with concomitant anterior expansion of expression domains of its targets, Hoxd10-13 genes, in the absence of ectopic Sonic hedgehog expression. Upstream regulation of Plzf in limb buds is currently unknown. We present here the first candidate Plzf cis-regulatory sequence.
authors.affiliation: Institute of Biology and Medical Genetics of the 1st Faculty of Medicine and General Teaching Hospital, Charles University in Prague, Praha, Czech Republic. frantisek.liska@lf1.cuni.cz
authors.forename: Frantisek
authors.initials: F
authors.lastname: Liska
authors.forename: Pavel
authors.initials: P
authors.lastname: Snajdr
authors.forename: Lucie
authors.initials: L
authors.lastname: Sedová
authors.forename: Ondrej
authors.initials: O
authors.lastname: Seda
authors.forename: Blanka
authors.initials: B
authors.lastname: Chylíková
authors.forename: Petra
authors.initials: P
authors.lastname: Slámová
authors.forename: Eliska
authors.initials: E
authors.lastname: Krejcí
authors.forename: David
authors.initials: D
authors.lastname: Sedmera
authors.forename: Milos
authors.initials: M
authors.lastname: Grim
authors.forename: Drahomíra
authors.initials: D
authors.lastname: Krenová
authors.forename: Vladimír
authors.initials: V
authors.lastname: Kren
mesh_terms: D000818:Animals; D001483:Base Sequence; D019521:Body Patterning; D017124:Conserved Sequence; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D015536:Down-Regulation / Q000235:genetics*; D004622:Embryo, Mammalian / Q000196:embryology / Q000378:metabolism; D017353:Gene Deletion; D018507:Gene Expression Regulation, Developmental; D007438:Introns / Q000235:genetics*; D018878:Limb Buds / Q000002:abnormalities* / Q000378:metabolism*; D017689:Polydactyly / Q000235:genetics / Q000378:metabolism*; D000075722:Promyelocytic Leukemia Zinc Finger Protein; D012333:RNA, Messenger / Q000235:genetics; D022661:RNA, Untranslated / Q000235:genetics*; D051381:Rats
chemical_list: D004268:DNA-Binding Proteins; D000075722:Promyelocytic Leukemia Zinc Finger Protein; D012333:RNA, Messenger; D022661:RNA, Untranslated; C534029:ZBTB16 protein, rat