Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “evolutionary conservation”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 1,207 records · Page 67Linked to original sources

Structure and expression of the mouse apolipoprotein C2 gene.

Three cDNA clones containing the mouse apolipoprotein C2 (Apoc2) gene were isolated from a mouse liver cDNA library. The inserts from two cDNA clones were 500 bp in size while the insert from the third clone was unexpectedly large, 962 bp. All three clones contained a single open reading frame encoding apoC2. The exon-intron structure of the mouse Apoc2 gene was determined by sequence analysis. Northern blotting and primer extension analysis of mouse RNA showed that the major liver transcript is 500 bp in size and is encoded by four exons. Transcripts for Apoc2 were found in fetal liver, adult liver, intestine, and peritoneal macrophages. The largest cDNA clone, mAPOC2c4, contained an additional 440 bp at the 5' end that are evolutionary conserved between man and mouse. These additional sequences are encoded by two exons located 5' to the major liver start site. Although the larger transcript could not be detected by Northern blot analysis, products resulting from an upstream transcription initiation site were detected in the liver using RT-PCR analysis. The sizes of the RT-PCR products are consistent with alternative splicing.

Amino Acid Sequence↗

Chromosomal localization of the gonadotropin-releasing hormone receptor gene to human chromosome 4q13.1-q21.1 and mouse chromosome 5.

The gonadotropin-releasing hormone receptor (GRHR) is a G-protein-coupled receptor on the cell surface of pituitary gonadotropes, where it serves to transduce signals from the extracellular ligand, the hypothalamic factor gonadotropin-releasing hormone, and to modulate the synthesis and secretion of luteinizing hormone and follicle-stimulating hormone. We have localized the GRHR gene to the q13.1-q21.1 region of the human chromosome 4 using mapping panels of human/rodent somatic cell hybrids containing different human chromosomes or different regions of human chromosome 4. Furthermore, using linkage analysis of single-strand conformational polymorphisms, the murine GRHR gene was localized to mouse chromosome 5, linked to the endogenous retroviral marker Pmv-11. This is consistent with te evolutionary conservation of homology between these two regions, as has been previously suggested from comparative mapping of several other loci. The localization of the GRHR gene may be useful in the study of disorders of reproduction.

Animals↗

The fibulin-1 gene (FBLN1) is located on human chromosome 22 and on mouse chromosome 15.

Fibulin-1 is a calcium-binding glycoprotein present in the extracellular matrix and in the serum. The gene coding for fibulin-1 (FBLN1) was located by in situ hybridization of 3H-labeled cDNA probes to human and mouse metaphase chromosomes. The gene was assigned to the q13.2-q13.3 region of human chromosome 22 and to the E-F band of mouse chromosome 15. This finding extends the evolutionary conservation between human chromosome 22 and mouse chromosome 15.

Animals↗

Genomic cloning of the mouse LDL receptor related protein/alpha 2-macroglobulin receptor gene.

The LDL receptor-related protein (LRP) or alpha 2-macroglobulin receptor (A2mr) is encoded by a 15-kb mRNA in mouse and human. Probes encompassing different regions of the mouse cDNA were used to isolate clones from a cosmid library of mouse strain 129. Four overlapping cosmids were used for restriction mapping and Southern blot analysis. This map and hybridization data obtained with oligonucleotide probes from the 5' and 3' ends of the Lrp cDNA demonstrated that the mouse gene is approximately 85 kb in size. The Lrp promoter region was sequenced and reveals strong evolutionary conservation of putative regulatory elements between mouse and human. The present study will facilitate detailed elucidation of the function of LRP in vivo.

Animals↗

The mouse gene Ptprf encoding the leukocyte common antigen-related molecule LAR: cloning, characterization, and chromosomal localization.

The human receptor-like protein tyrosine phosphatase leukocyte common antigen-related molecule (LAR; gene symbol PTPRF) closely resembles cell adhesion molecules, which suggests that it may be involved in the regulation of phosphotyrosine levels through cell-cell or cell-matrix interactions. To obtain a better understanding of LAR function, we have characterized the mouse Ptprf gene as a first step toward site-directed mutagenesis studies in vitro and in vivo. The cytoplasmic region of the mouse LAR (mLAR) protein is encoded by 11 exons that span only 4.5 kb of genomic DNA. Compared to the known exon-intron structures of other mammalian receptor-like protein tyrosine phosphatase genes, such as Ptpra (encoding LRP) and Ptprc (coding for Ly-5), the Ptprf gene part encoding the cytoplasmic region of mLAR contains not only smaller, but also fewer introns. Sequence analysis of both phosphatase domains of mLAR and its homologs MPTP delta and mRPTP sigma revealed a higher evolutionary conservation of the second, C-terminal domain in comparison to the first domain. Fluorescence in situ hybridization was used to map the Ptprf gene to region C6-D1 on mouse chromosome 4.

Amino Acid Sequence↗

Structure and chromosomal localization of the human stromal cell-derived factor 1 (SDF1) gene.

Stromal cell-derived factors 1 alpha and 1 beta are small cytokines belonging to the intercrine CXC subfamily and originally isolated from a murine bone-marrow stroma cell line by the signal sequence trap method. cDNA and genomic clones of human SDF1 alpha and SDF1 beta (SDF1A and SDF1B) were isolated and characterized. cDNAs of SDF1 alpha and SDF1 beta encode proteins of 89 and 93 amino acids, respectively. SDF1 alpha and SDF1 beta sequences are more than 92% identical to those of the human counterparts. The genomic structure of the SDF1 gene revealed that human SDF1 alpha and SDF1 beta are encoded by a single gene and arise by alternative splicing. SDF1 alpha and SDF1 beta are encoded by 3 and 4 exons, respectively. Ubiquitous expression of the SDF1 gene, except in blood cells, was consistent with the presence of the GC-rich sequence in the 5'-flanking region of the SDF1 gene, as is often the case in the "housekeeping" genes. Although genes encoding other members of the intercrine family are localized on chromosome 4q or 17q, the human SDF1 gene was mapped to chromosome 10q by fluorescence in situ hybridization. Strong evolutionary conservation and unique chromosomal localization of the SDF1 gene suggest that SDF1 alpha and SDF1 beta may have important functions distinct from those of other members of the intercrine family.

Amino Acid Sequence↗

CpG islands in human ZFX and ZFY and mouse Zfx genes: sequence similarities and methylation differences.

The human ZFX, human ZFY, and mouse Zfx genes have CpG islands near their 5; ends. These islands are typical in that they span about 1.5 kb, contain transcription initiation sites, and encompass some 5' untranslated exons and introns. However, comparitive nucleotide sequencing of these human and mouse islands provided evidence of evolutionary conservation to a degree unprecedented among mammalian 5' CpG islands. In one stretch of 165 nucleotides containing 19 CpGs, mouse Zfx and human ZFX are identical to each other and differ from human ZFY at only 9 nucleotides. In contrast, we found no evidence of homologous CpG islands in the mouse Zfy genes, whose transcription is more circumscribed than that of human ZFX, human ZFY, and mouse Zfx. Using the isoschizomers HpaII and MspI to examine a highly conserved segment of the ZFX CpG island, we detected methylation on inactive mouse X chromosomes but not on inactive human X chromosomes. These observations parallel the previous findings that mouse Zfx undergoes X inactivation while human ZFX escapes it.

Animals↗

The human interleukin-11 receptor alpha gene (IL11RA): genomic organization and chromosome mapping.

The high-affinity receptor for interleukin-11 (IL-11) is composed of two subunits, IL-11 receptor alpha chain (IL-11R alpha) and gp130, the common subunit of the interleukin-6 (IL-6), ciliary neurotrophic factor (CNTF), leukemia inhibitory factor, and oncostatin M receptors. The IL-11 receptor-specific alpha chain shares homologies with the alpha chain of the CNTF and IL-6 receptors. We isolated and characterized genomic DNA clones encompassing the entire coding sequence of the IL-11R alpha cDNA. The exon-intron organization of the IL-11R gene (HGMW-approved symbol IL11RA) is consistent with the predicted structure of the different domains of the IL-11R alpha protein, confirming evolutionary conservation at the level of gene organization among the hematopoietic cytokine receptor family. The IL-11R gene has been assigned to chromosome 9 band p13 by in situ hybridization using human IL-11R alpha cDNA as a probe. The fact that the ciliary neurotrophic factor (CNTFR) gene has recently been localized on this same band and the conserved genomic structure between IL-11R and CNTFR suggest that they may have evolved from a common ancestor.

Base Sequence↗

Cloning, mapping, and in vivo localization of a human member of the PKCI-1 protein family (PRKCNH1).

We report here the complete cDNA sequence, genomic mapping, and immunolocalization of the first human member of the protein kinase C inhibitor (PKCI-1) gene family. The predicted human protein (hPKCI-1) is 96% identical to bovine and 53% identical to maize members, indicating the great evolutionary conservation of this protein family. The hPKCI-1 gene (HGMV-approved symbol PRKCNH1) maps to human chromosome 5q31.2 by fluorescence in situ hybridization. Indirect immunofluorescence shows that hPKCI-1 localizes to cytoskeletal structures in the cytoplasm of a human fibroblast cell line and is largely excluded from the nucleus. The cytoplasmic localization of hPKCI-1 is consistent with a postulated role in mediating a membrane-derived signal in response to ionizing radiation.

Amino Acid Sequence↗

Molecular cloning of a novel human gene (D11S4896E) at chromosomal region 11p15.5.

A novel gene (GOK) has been cloned from human chromosome region 11p15.5 that is believed to contain a gene or genes associated with a number of pediatric malignancies, including Wilms tumor. A 4-kb cDNA has been cloned and it encodes a predicted protein of approximately 84 kDa that could be translated in vitro. Computer analysis predicted that the protein had a signal peptide and may contain a transmembrane helix. Restriction mapping by pulsed-field electrophoresis indicates that GOK is located 1.7 kb telomeric of RRM1, and both genes are transcribed in the same direction. GOK displays high evolutionary conservation: cloning and partial sequencing of a mouse genomic clone revealed 90% identity with the human gene at both the nucleotide and the predicted amino acid levels.

Amino Acid Sequence↗

SMT3A, a human homologue of the S. cerevisiae SMT3 gene, maps to chromosome 21qter and defines a novel gene family.

cDNA selection was used to isolate coding sequences from cosmids mapping to the gene-rich telomeric region of human chromosome 21q. A novel cDNA, termed SMT3A, was isolated and mapped between the loci PFKL and D21S171, about 2.2 Mb proximal to the telomere. The predicted protein of 103 amino acids appears to be a homologue of the Saccharomyces cerevisiae SMT3 protein, whose gene was previously isolated as a suppressor of mutations in the MIF2 gene. The yeast MIF2 gene encodes an essential centromeric protein and shows homology to mammalian CENP-C, an integral component of active kinetochores. SMT3A was found to be highly homologous to two other recently isolated human genes, suggesting the presence of a new gene family. Homologous sequences were also found in protozoa, metazoa, and plants. Moreover, all predicted proteins show significant homology to ubiquitin. The proposed role of yeast SMT3 as centromeric protein and the strong evolutionary conservation of the SMT3A gene suggest an involvement of the encoded protein in the function and/or structure of the eukaryotic kinetochore.

Amino Acid Sequence↗

Genomic organization and molecular characterization of a gene encoding HsPXF, a human peroxisomal farnesylated protein.

A protein modification essential for the cellular sorting of many biologically relevant proteins is the covalent attachment of prenyl lipids by specific transferases. Isoprenylation is known to render protein domains hydrophobic, thereby facilitating the interaction with lipid bilayers and/or membrane proteins. The target for the modification with farnesyl groups is the COOH-terminal sequence CaaX. Among the variety of farnesylated proteins the only one reported so far to be located to peroxisomes is the 37-kDa peroxisomal farnesylated hamster protein PxF. Recently we published data on the cDNA of the human gene HK33 (A. Braun et al., 1994, Gene 146: 291-295), which was revealed to be the human ortholog of PxF and was consequently renamed HsPXF. The genomic structure, molecular characterization, and evolutionary conservation of HsPXF are described herein. The exact location of the gene was defined as chromosome 1q22. The gene spans a region of approximately 9 kb, containing eight exons and seven introns. The 5' upstream region showed two potential Sp1-binding sites and an Alu repetitive sequence. Luciferase reporter activating capacity confirmed the presumed promoter activity of this region. On the transcriptional level, we detected four splice variants originating either from exon skipping or from alternative splicing events. For the HsPXF protein, a carboxyterminal farnesylation at cysteine residues was demonstrated. Through the use of HsPXF-specific antibodies, the protein was shown to be attached to the outer surface of peroxisomes. This localization together with the similarity to a peroxisomal assembly protein from Saccharomyces cerevisiae suggests HsPXF is involved in the process of peroxisomal biogenesis or assembly.

Amino Acid Sequence↗

Identification and characterization of the human ortholog of rat STXBP1, a protein implicated in vesicle trafficking and neurotransmitter release.

In a screen designed to identify genes expressed preferentially in retina, we identified a cDNA encoding the human ortholog of rat STXBP1 (n-Sec1, Munc-18-1, rbSec1), a protein implicated in vesicle trafficking and neurotransmitter release. This protein also has similarity to Drosophila Rop (65% aa identity) and Caenorhabditis elegans UNC-18 (58% aa identity). The major human cDNA encodes a protein of 594 amino acids which has 100 % amino acid identity with its rat and murine counterparts. Additionally, there is an alternative splice form in humans, arising from the inclusion of an additional exon, which encodes a protein of 603 amino acids and is also 100% identical to the corresponding rat isoform. We found expression of the shorter cDNA in all tissues and cell lines we examined with highest levels in retina and cerebellum. By RT-PCR analysis, we found expression of the longer cDNA in neural tissues only. We mapped the structural gene to 9q34.1, a region without obvious candidate phenotypes. However, due to its evolutionary conservation and abundant expression in retina and brain, STXBP1 should be considered a candidate gene for retinal and/or neural disorders mapping to 9q34.1.

Amino Acid Sequence↗

Molecular cloning and characterization of the human p44 mitogen-activated protein kinase gene.

The complete genomic structure of the human p44(mapk) gene (HMGW-approved symbol PRKM3) has been determined. The gene covers 9 kb and is composed of nine exons and eight introns. This structure is identical to the previously reported mouse p44(mapk) gene, indicating a high degree of evolutionary conservation. A sequence differing by one nucleotide from the consensus TATA box is present 132 positions upstream of the main transcription initiation point. This point has been located 415 nucleotides upstream of the translation initiation codon ATG and perfectly meets the consensus criteria for an initiator element (Inr). Multiple consensus sequences for factors that regulate either basal transcription or gene expression during cell differentiation and proliferation can be found in the putative promoter region. Some of them, such as several G/C boxes located downstream from the transcription initiation point, are also present in the homologous mouse gene, where they were shown to be functional.

5' Untranslated Regions↗

The murine fem1 gene family: homologs of the Caenorhabditis elegans sex-determination protein FEM-1.

The pathway controlling sex determination in the nematode Caenorhabditis elegans is a model for the genetic control of cell-fate determination. We report here the cloning and characterization of a new mouse gene family with homology to FEM-1, a signal-transducing regulator in the C. elegans sex-determination pathway. This gene family consists of two known members, designated Fem1a and Fem1b. The highest degree of homology between the two mouse proteins and the nematode protein is in a domain that encodes seven sequential ANK repeats. The Fem1a gene localizes to chromosome 17 and is highly expressed in adult heart and skeletal muscle. The Fem1b gene localizes to chromosome 9 and is highly expressed in adult testis. Both genes are expressed during embryogenesis. The existence of FEM-1 homologs in the mouse raises the possibility that evolutionary conservation of ancient FEM-1 signaling interactions may play a role in vertebrate cell-fate determination.

Amino Acid Sequence↗

DXS6673E encodes a predominantly nuclear protein, and its mouse ortholog DXHXS6673E is alternatively spliced in a developmental- and tissue-specific manner.

DXS6673E is a candidate gene for nonspecific X-linked mental retardation and encodes a novel Zn-finger protein. The ortholog murine gene DXHXS6673E in XC-D was isolated and characterized. It is ubiquitously expressed in all embryonic stages and adult tissues. Two different transcription start sites exist that result in two major transcripts of 6055 and 5352 nucleotides, each composed of 25 exons. Exon 1A is tissue specific, whereas exon 1B is transcribed constitutively. Both variants are translated into the same 1370-amino-acid protein. Transcripts are subject to alternative splicing at the 5'-end. Some of the isoforms are developmental stage and tissue specific. Among them, one was present only in embryos and adult brain. Sequence analysis demonstrated evolutionary conservation down to the arthropods and defined several conserved protein motifs. Subcellular localization studies with green fluorescent protein as a reporter showed that DXS6673E is predominantly located in the nucleus due to several functional nuclear localization signals. Three distinct protein distribution patterns in COS-7 cells could be identified.

Alternative Splicing↗

Structure, expression, and chromosomal localization of the mouse Atox1 gene.

Copper trafficking in eukaryotes involves small proteins termed metallochaperones, which mediate copper delivery to specific intracellular sites. Previous studies in yeast and human cell lines have suggested that Atox1 plays a critical role in copper delivery to the secretory pathway. In the present study, a mouse Atox1 (mAtox1) cDNA was cloned and shown to encode an open reading frame with 85% amino acid identity to human Atox1. RNA blot analysis revealed that mAtox1 was expressed as a single transcript in multiple tissues, and immunoblotting indicated that the relative abundance of mAtox1 mRNA directly correlated with mAtox1 protein. Analysis of the mAtox1 gene locus revealed a genomic structure with four exons encompassing a total of 14.5 kb. RFLP and haplotype analyses indicated that the mAtox1 locus was tightly linked to the Trhr and D15Bir7 loci on mouse chromosome 15. Taken together, these data reveal marked evolutionary conservation of Atox1 structure and provide a genomic organization and localization that will aid in the genetic deciphering of the molecular role of this protein in copper homeostasis.

Amino Acid Sequence↗

Cloning of the murine Krit1 cDNA reveals novel mammalian 5' coding exons.

Human KRIT1 (Krev interaction trapped 1), a defective gene product in cerebral cavernous malformation, was cloned from a HeLa cell cDNA library by virtue of its interaction with Krev/rap1A, a small ras-family GTPase. We have now characterized the full-length cDNA for the murine orthologue that encodes a predicted protein of 736 amino acids, 207 amino acids longer than the previously reported human protein. 5' Rapid amplification of cDNA ends analysis of mouse mRNA demonstrated a single transcriptional start site. The putative initiator codon was found within a context that conformed well to the Kozak consensus sequence and was preceded by an in-frame termination codon. BLAST analysis revealed that conceptual translation of a fragment of human genomic DNA upstream of the 5' end of the reported KRIT1 coding sequence predicts extension of the human open reading frame by 207 codons with 95% amino acid identity between the novel putative human and murine amino termini. This block of coding sequence was divided among four exons that are flanked by consensus splice site sequences. The extreme evolutionary conservation of this region, including a putative nuclear localization signal, indicates functional importance. These data have immediate relevance to mutation screening efforts in cerebral cavernous malformation and may contribute to our understanding of the normal biology of KRIT1 and the pathogenesis of this disorder.

Amino Acid Sequence↗