RESUMEN
Zinc-finger (ZF) proteins are widely distributed. The current study reports isolation, cloning and characterization of a novel ZF gene, RLZF-Y. Total RNA from rat lung was reverse transcribed. The 5' and 3' ends were isolated by rapid amplification of cDNA ends (RACE) using primers derived from a previously isolated partial clone. RACE products of 1.5 and 1.1 kb were cloned and sequenced. Identical overlapping sequence of 70 base pairs confirmed representation of the same cDNA approximately 2.5 kb in length. Probes derived from both 5' RACE and 3' RACE products independently hybridized to a 2.5 kb mRNA from rat lung. RLZF-Y mRNA is expressed in lung, brain, heart and kidney; expression is low in liver. Predicted amino acid sequence analysis defined three regions of similarity to known C2H2 ZF proteins: a region containing seven ZF structures characteristic of the Krüppel-like subfamily of ZF genes; a region with sequence similarity to the Krüppel-associated box A (KRAB-A) domain at the amino end; an amino-terminal leucine-rich region (LeR) adjacent to KRAB-A. The presence of KRAB-A and the adjacent LeR implies RLZF-Y protein may function as a transcriptional repressor.
Asunto(s)
Proteínas de Unión al ADN/genética , Ratas/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Clonación Molecular , Cartilla de ADN , Proteínas de Unión al ADN/biosíntesis , Proteínas de Unión al ADN/química , Humanos , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , ARN Mensajero/genética , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/química , Alineación de Secuencia , Homología de Secuencia de Aminoácido , Transcripción Genética , Dedos de Zinc/genéticaRESUMEN
Control of alveolar cell growth and differentiation after pneumonectomy likely involves changes in expression of regulatory genes, including those encoding zinc finger (ZF) proteins. To explore this premise, total RNA from the lungs of control and pneumonectomized rats was reverse transcribed; PCRs were performed with degenerate primers corresponding to amino acid sequences HTGEKP and CPECGK(N), which are evolutionarily conserved among ZF genes. Reaction products corresponding to three and four ZF units were isolated and cloned. Sixteen clones were sequenced and found to represent rat lung ZF genes: six clones were highly similar or identical to known ZF genes and ten clones showed lower homology to known ZF genes and thus appear to represent new members of the ZF family. Northern analysis demonstrated differential expression of some ZF genes after pneumonectomy. Thus a PCR-based strategy with primers derived from evolutionarily conserved ZF protein sequences efficiently identifies ZF genes expressed in lung, some of which may play a role in cellular growth and differentiation.