ABSTRACT
An improved linkage map for rat Chromosome (Chr) 10 with two F2 populations was constructed. Thirty new microsatellite markers were generated from a Chr 10-specific, small-insert genomic library and mapped to rat Chr 10. Among them were the rat homologs for the mouse gene for light and heavy chains of myeloperoxidase and human neurofibromatosis 1. Eight newly generated markers (D10Mco62, D10Mco63, D10Mco64, D10Mco65, D10Mco67, D10Mco68, D10Mco70, and D10Mco74) were mapped to the region of the rat Chr 10 blood pressure QTL. The availability of such markers may be instrumental in the search for genes responsible for the hypertension.
Subject(s)
Genetic Linkage , Microsatellite Repeats , Animals , Chromosome Mapping , Molecular Sequence Data , Rats , Rats, Inbred Lew , Rats, Inbred SHR , Rats, Inbred Strains , Rats, WistarABSTRACT
Fifty-eight new anonymous simple sequence repeats (SSR) were generated and mapped to various rat chromosomes. Among them two genes (rat homologs for human cadherin-14 and mouse fibroblast growth factor-related protein) were mapped on Chromosomes (Chrs) 2 and 11 respectively. The majority of markers were generated from a small insert genomic library specific to Chr 11, 13, 14, and 15. Twenty new markers were mapped to Chr 13, which is known to contain a blood pressure quantitative trait locus (QTL). Several approaches to obtain microsatellite markers are described. The protocols and newly generated markers should be useful for ongoing rat genome project.
Subject(s)
Chromosome Mapping/methods , Genetic Linkage , Microsatellite Repeats , Rats/genetics , Animals , Base Sequence , DNA Primers/genetics , Humans , Mice , Molecular Sequence Data , Rats, Inbred StrainsSubject(s)
Chromosome Mapping , Genetic Linkage , Microsatellite Repeats , Animals , Base Sequence , Chromosomes , DNA, Complementary , Molecular Sequence Data , RatsABSTRACT
Human ATP1AL1 and corresponding genes of other mammals encode the catalytic alpha subunit of a non-gastric ouabain-sensitive H,K-ATPases, the ion pump presumably involved in maintenance of potassium homeostasis. The tissue specificity of the expression of these genes in different species has not been analyzed in detail. Here we report comparative RT-PCR screening of mouse, rat, rabbit, human, and dog tissues. Significant expression levels were observed in the skin, kidney and distal colon of all species (with the exception of the human colon). Analysis of rat urogenital organs also revealed strong expression in coagulating and preputial glands. Relatively lower expression levels were detected in many other tissues including brain, placenta and lung. In rabbit brain the expression was found to be specific to choroid plexus and cortex. Prominent similarity of tissue-specific expression patterns indicates that animal and human non-gastric H,K-ATPases are indeed products of homologous genes. This is also consistent with the high sequence similarity of non-gastric H,K-ATPases (including partial sequences of hitherto unknown cDNAs for mouse and dog proteins).
Subject(s)
H(+)-K(+)-Exchanging ATPase/genetics , Ouabain/metabolism , Amino Acid Sequence , Animals , Brain/enzymology , Catalytic Domain , Colon/enzymology , Dogs , Gene Expression , H(+)-K(+)-Exchanging ATPase/metabolism , Humans , Kidney/enzymology , Mice , Molecular Sequence Data , Placenta/enzymology , Rabbits , Rats , Rats, Sprague-Dawley , Reverse Transcriptase Polymerase Chain Reaction , Sequence Homology, Amino Acid , Skin/enzymology , Tissue Distribution , Urogenital System/enzymologyABSTRACT
The human ATP1AL1 gene belongs to the family of Na,K-ATPase and H,K-ATPase (X,K-ATPases) genes. It encodes a catalytic subunit of hitherto unknown human ouabain-sensitive H,K-ATPase that represents a novel third group of X,K-ATPases distinct from the known Na,K-ATPase and gastric H,K-ATPase. Cloning of the ATP1AL1 gene is described in this report. The exon-intron structure of ATP1AL1 was found to be very similar to that of related genes. It contains 23 exons and spans approximately 32 kb of genomic DNA. All ATP1AL1 exons and 12 of its 22 introns were entirely sequenced. A total of nine Alu repeats were identified in introns. The transcription initiation site was mapped 187 bp upstream of the ATG initiation codon by primer extension and S1 nuclease protection analyses of RNA from human skin and colon. Sequence analysis of the 5'-flanking region (1.48 kb) revealed numerous potential binding sites for transcription factors Sp1 and AP2 and one putative NF-kappa B binding site. The 0.85-kb region from position -484 (5'-flanking region) to position +369 (intron 1) meets the structural criteria of a CpG island. It is suggested that the ATP1AL1 gene contains two poly(A) addition sites that may function in a tissue-specific manner.
Subject(s)
Genes/genetics , H(+)-K(+)-Exchanging ATPase/genetics , Sodium-Potassium-Exchanging ATPase/genetics , Amino Acid Sequence , Base Sequence , Cloning, Molecular , Colon/chemistry , Exons/genetics , Humans , Introns/genetics , Kidney/chemistry , Molecular Sequence Data , Ouabain , RNA, Messenger/analysis , Repetitive Sequences, Nucleic Acid/genetics , Restriction Mapping , Sequence Alignment , Sequence Analysis, DNA , Skin/chemistry , Transcription, Genetic/geneticsABSTRACT
The cDNA for ATP1AL1--the fifth member of the human Na,K-/H,K-ATPase gene family--was cloned and sequenced. The deduced primary ATP1AL1 translation product is 1,039 amino acids in length and has Mr of 114,543. The encoded protein has all of the structural features common to known catalytic subunits of P-type membrane ion-transporting ATPases and is equally distant (63-64% of identity) from the Na,K-ATPase isoforms and the gastric H,K-ATPase. The ATP1AL1 encoded protein was proposed to represent a new separate group within the family of human potassium-dependent ion pumps.
Subject(s)
Adenosine Triphosphatases/classification , H(+)-K(+)-Exchanging ATPase/genetics , Multigene Family/genetics , Sodium-Potassium-Exchanging ATPase/genetics , Adenosine Triphosphatases/genetics , Amino Acid Sequence , Base Sequence , DNA, Complementary/genetics , H(+)-K(+)-Exchanging ATPase/chemistry , H(+)-K(+)-Exchanging ATPase/classification , Humans , Ion Pumps/metabolism , Kidney/enzymology , Molecular Sequence Data , Potassium/metabolism , Sequence Homology, Amino Acid , Skin/enzymology , Sodium-Potassium-Exchanging ATPase/chemistry , Sodium-Potassium-Exchanging ATPase/classification , Tissue DistributionABSTRACT
The multigene family of human Na,K-ATPase is composed of 5 alpha-subunit genes, 3 of which were shown to encode the functionally active alpha 1, alpha 2 and alpha 3 isoforms of the catalytic subunits. This report describes the isolation, mapping and partial sequencing of the fourth gene (ATP1AL1) that was demonstrated here to be functionally active and expressed in human brain and kidney. Limited DNA sequencing of the ATP1AL1 exons allowed one to suggest that the gene probably encodes a new ion transport ATPase rather than an isoform of the Na,K-ATPase or the closely related H,K-ATPase.
Subject(s)
Adenosine Triphosphatases/genetics , Multigene Family , Sodium-Potassium-Exchanging ATPase/genetics , Transcription, Genetic , Adenosine Triphosphatases/metabolism , Amino Acid Sequence , Base Sequence , Biological Transport , Brain/enzymology , Electrophoresis, Agar Gel , Exons , Humans , Introns , Kidney/enzymology , Molecular Sequence Data , Oligonucleotide Probes , Polymerase Chain Reaction , Sequence Homology, Nucleic AcidABSTRACT
Intra-individual tissue-specific restriction fragment length polymorphism (RFLP) has been demonstrated in DNA isolated from different mammalian tissues using cDNAs of alpha- and beta-subunits of Na+,K+-ATPase as hybridization probes. We propose that the RFLPs could result from gene rearrangements in the gene loci for the alpha- and beta-subunits of Na+,K+-ATPase. The changes in restriction patterns have been shown to occur during embryonic development and tumor formation. In addition, the tissue specificity of the expression of different genes of the family of Na+,K+-ATPase genes and their low expression in tumor cells have been demonstrated.
