scholarly journals ERV3 and Related Sequences in Humans: Structure and RNA Expression

2005 ◽  
Vol 79 (14) ◽  
pp. 9270-9284 ◽  
Author(s):  
Ann-Catrin Andersson ◽  
Zhihong Yun ◽  
Göran O. Sperber ◽  
Erik Larsson ◽  
Jonas Blomberg
Keyword(s):  
Adrenal Gland ◽  
Binding Sites ◽  
Adrenal Glands ◽  
Open Reading Frame ◽  
Rna Expression ◽  
Human Tissues ◽  
Sebaceous Glands ◽  
Reading Frame ◽  
Expression Control

ABSTRACT The ERV3 locus at chromosome 7q11 is a much studied human endogenous retroviral (HERV) sequence, owing to an env open reading frame (ORF) and placental RNA and protein expression. An analysis of the human genome demonstrated that ERV3 is one of a group of 41 highly related elements (ERV3-like HERVs) which use proline, isoleucine, or arginine tRNA in their primer binding sites. In addition to elements closely related to ERV3, the group included the previously known retinoic acid-inducible element, RRHERVI, also referred to as HERV15, but was separate from the related HERV-E elements. The ERV3-like elements are defective. The only element with an ORF among gag, pro, pol, and env genes was the env ORF of the original ERV3 locus. A search in dbEST revealed ERV3 RNA expression in placenta, skin, carcinoid tumor, and adrenal glands. Expression was also studied with newly developed real-time quantitative PCRs (QPCR) of ERV3 and HERV-E(4-1) env sequences. Results from a novel histone 3.3 RNA QPCR result served as the expression control. QPCR results for ERV3 were compatible with previously published results, with a stronger expression in adrenal gland and placenta than in 15 other human tissues. The expression of the envelope (env) of ERV3 at chromosome 7q11 was also studied by using stringent in situ hybridization. Expression was found in corpus luteum, testis, adrenal gland, Hassal's bodies in thymus, brown fat, pituitary gland, and epithelium of the lung. We conclude that ERV3 env is most strongly expressed in adrenal and sebaceous glands as well as in placenta.

Molecular cloning, characterization, and distribution of the gerbil angiotensin II AT2 receptor

2003 ◽  
Vol 285 (6) ◽  
pp. R1373-R1383 ◽  
Author(s):  
Kwang-Lae Hoe ◽  
Ines Armando ◽  
Gustavo Baiardi ◽  
Taduru Sreenath ◽  
Ashok Kulkarni ◽  
...  
Keyword(s):  
Open Reading Frame ◽  
Late Gestation ◽  
At2 Receptor ◽  
Ang Ii ◽  
Amino Acid Residues ◽  
Receptor Ligands ◽  
Reading Frame ◽  
Receptor Mrna ◽  
Dental Papilla

We isolated a cDNA clone encoding the gerbil AT2 receptor (gAT2) gene from a gerbil adrenal gland cDNA library. The full-length cDNA contains a 1,089-bp open reading frame encoding 363 amino acid residues with 90.9, 96.1, and 95.6% identity with the human (hAT2), rat (rAT2), and mouse AT2 (mAT2) receptors, respectively. There are at least seven nonconserved amino acids in the NH2-terminal domain and in positions Val196, Val217, and Met293, important for angiotensin (ANG) II but not for CGP-42112 binding. Displacement studies in adrenal sections revealed that affinity of the gAT2 receptor was 10-20 times lower for ANG II, ANG III, and PD-123319 than was affinity of the rAT2 receptor. The affinity of each receptor remained the same for CGP-42112. When transfected into COS-7 cells, the gAT2 receptor shows affinity for ANG II that is three times lower than that shown by the hAT2 receptor, whereas affinities for ANG III and the AT2 receptor ligands CGP-42112 and PD-123319 were similar. Autoradiography in sections of the gerbil head showed higher binding in muscles, retina, skin, and molars at embryonic day 19 than at 1 wk of age. In situ hybridization and emulsion autoradiography revealed that at embryonic day 19 the gAT2 receptor mRNA was highly localized to the base of the dental papilla of maxillary and mandibular molars. Our results suggest selective growth-related functions in late gestation and early postnatal periods for the gAT2 receptor and provide an essential basis for future mutagenesis studies to further define structural requirements for agonist binding.

scholarly journals Identification of the miaB Gene, Involved in Methylthiolation of Isopentenylated A37 Derivatives in the tRNA of Salmonella typhimurium andEscherichia coli

1999 ◽  
Vol 181 (23) ◽  
pp. 7256-7265 ◽  
Author(s):  
Birgitta Esberg ◽  
Hon-Chiu Eastwood Leung ◽  
Ho-Ching Tiffany Tsui ◽  
Glenn R. Björk ◽  
Malcolm E. Winkler
Keyword(s):  
Escherichia Coli ◽  
Salmonella Typhimurium ◽  
Binding Site ◽  
Binding Sites ◽  
Open Reading Frame ◽  
Iron Binding ◽  
Reading Frame ◽  
Conserved Motif ◽  
E Coli ◽  
Weak Similarity

ABSTRACT The tRNA of the miaB2508::Tn10dCm mutant of Salmonella typhimurium is deficient in the methylthio group of the modified nucleosideN 6-(4-hydroxyisopentenyl)-2-methylthioadenosine (ms2io6A37). By sequencing, we found that the Tn10dCm of this strain had been inserted into thef474 (yleA) open reading frame, which is located close to the nag locus in both S. typhimurium and Escherichia coli. By complementation of the miaB2508::Tn10dCm mutation with a minimal subcloned f474 fragment, we showed thatf474 could be identified as the miaB gene, which is transcribed in the counterclockwise direction on the bacterial chromosome. Transcriptional studies revealed two promoters upstream ofmiaB in E. coli and S. typhimurium. A Rho-independent terminator was identified downstream of themiaB gene, at which the majority (96%) of themiaB transcripts terminate in E. coli, showing that the miaB gene is part of a monocistronic operon. A highly conserved motif with three cysteine residues was present in MiaB. This motif resembles iron-binding sites in other proteins. Only a weak similarity to an AdoMet-binding site was found, favoring the idea that the MiaB protein is involved in the thiolation step and not in the methylating reaction of ms2i(o)6A37 formation.

The End of the LINE?: Lack of Recent L1 Activity in a Group of South American Rodents

Genetics ◽  
2000 ◽  
Vol 154 (4) ◽  
pp. 1809-1817
Author(s):  
N Carol Casavant ◽  
LuAnn Scott ◽  
Michael A Cantrell ◽  
Lara E Wiggins ◽  
Robert J Baker ◽  
...  
Keyword(s):  
Southern Blot Analysis ◽  
Open Reading Frame ◽  
Mammalian Host ◽  
South American ◽  
Reading Frame ◽  
Species Specific ◽  
Long Interspersed Nuclear Element ◽  
Intact Open Reading Frame ◽  
Genomic Southern Blot Analysis

Abstract L1s (LINE-1: Long Interspersed Nuclear Element 1) are present in all mammals examined to date. They occur in both placental mammals and marsupials and thus are thought to have been present in the genome prior to the mammalian radiation. This unusual conservation of a transposable element family for over 100 million years has led to speculation that these elements provide an advantage to the genomes they inhabit. We have recently identified a group of South American rodents, including rice rats (Oryzomys), in which L1s appear to be quiescent or extinct. Several observations support this conclusion. First, genomic Southern blot analysis fails to reveal genus-specific bands in Oryzomys. Second, we were unable to find recently inserted elements. Procedures to enrich for young elements did not yield any with an intact open reading frame for reverse transcriptase; all elements isolated had numerous insertions, deletions, and stop codons. Phylogenetic analysis failed to yield species-specific clusters among the L1 elements isolated, and all Oryzomys sequences had numerous private mutations. Finally, in situ hybridization of L1 to Oryzomys chromosomes failed to reveal the characteristic L1 distribution in Oryzomys with either a homologous or heterologous probe. Thus, Oryzomys is a viable candidate for L1 extinction from a mammalian host.

scholarly journals Yellow Lupine Cyclophilin Transcripts Are Highly Accumulated in the Nodule Meristem Zone

2001 ◽  
Vol 14 (12) ◽  
pp. 1384-1394 ◽  
Author(s):  
Katarzyna Nuc ◽  
Przemysław Nuc ◽  
Ryszard Słomski
Keyword(s):  
In Situ Hybridization ◽  
Genomic Dna ◽  
Root Nodules ◽  
Open Reading Frame ◽  
Intronless Gene ◽  
Reading Frame ◽  
Gene Coding ◽  
Meristem Zone ◽  
Yellow Lupine

Cyclophilin (CyP) is one of the enzymes that act as peptidylprolyl cis-trans isomerases (EC 5.2.1.8). The cDNA and an intronless gene coding for cytosolic CyP have been isolated from yellow lupine. The deduced amino acid sequence of the characterized open reading frame shows approximately 80% homology with cytosolic CyP from other organisms. Southern blots of genomic DNA indicate that there is a small family of genes for CyP-related genes in the yellow lupine genome. RNA blot analyses demonstrate that CyP genes are expressed in all plant organs. The amount of CyP transcripts is dramatically increased in root nodules. In situ hybridization experiments indicate that CyP transcripts are localized mainly in meristematic tissues, with the highest level observed in the nodule meristem zone. The promoter of the sequenced gene contains 5′ AAAGAT 3′ and AT-rich motifs that are characteristic for some nodulin promoters.

Complete sequence of rabbit kidney aminopeptidase N and mRNA localization in rabbit kidney by in situ hybridization

1993 ◽  
Vol 71 (5-6) ◽  
pp. 278-287 ◽  
Author(s):  
Xiao-Feng Yang ◽  
Pierre Emmanuel Milhiet ◽  
Florence Gaudoux ◽  
Philippe Crine ◽  
Guy Boileau
Keyword(s):  
In Situ Hybridization ◽  
Transmembrane Domain ◽  
Complete Sequence ◽  
Aminopeptidase N ◽  
Full Length ◽  
Open Reading Frame ◽  
Rabbit Kidney ◽  
Mrna Levels ◽  
Reading Frame

We have isolated and sequenced a full-length cDNA for rabbit kidney aminopeptidase N (APN). The 3-kilobase cDNA contains 12 nucleotides of the 5′ noncoding region, a 2898 nucleotide long open reading frame, and 113 nucleotides of the 3′ untranslated region. The open reading frame encodes a type II membrane protein of 966 amino acid residues composed of a 10 residue NH2-terminal cytosolic domain, a 23 residue transmembrane domain, and a large 933 residue ectodomain that contains the active site. Rabbit APN has eight potential N-glycosylation sites and seven cysteine residues, one of which is located in the transmembrane domain. Computer analysis showed that the enzymes from human, rat, and rabbit were highly conserved, except for the stalk region immediately downstream from the transmembrane domain. Using in situ hybridization techniques we showed that in rabbit kidney, APN mRNA is present in proximal tubules but not in glomeruli, which corresponds to the localization of the protein observed by immunohistochemistry. Taken together, our results strongly suggest that the expression of APN in kidney is modulated at mRNA levels and not at translational and (or) posttranslational levels.Key words: aminopeptidase N, rabbit kidney, full-length sequence, in situ hybridization.

scholarly journals Translational Repression of the RpoS Antiadapter IraD by CsrA Is Mediated via Translational Coupling to a Short Upstream Open Reading Frame

mBio ◽  
2017 ◽  
Vol 8 (4) ◽  
Author(s):  
Hongmarn Park ◽  
Louise C. McGibbon ◽  
Anastasia H. Potts ◽  
Helen Yakhnin ◽  
Tony Romeo ◽  
...  
Keyword(s):  
Stationary Phase ◽  
Translation Initiation ◽  
Binding Sites ◽  
Sigma Factor ◽  
Rna Structure ◽  
Rna Binding ◽  
Open Reading Frame ◽  
Translational Coupling ◽  
Reading Frame ◽  
Ribosome Binding

ABSTRACT CsrA is a global regulatory RNA binding protein that has important roles in regulating carbon metabolism, motility, biofilm formation, and numerous other cellular processes. IraD functions as an antiadapter protein that inhibits RssB-mediated degradation of RpoS, the general stress response and stationary-phase sigma factor of Escherichia coli . Here we identified a novel mechanism in which CsrA represses iraD translation via translational coupling. Expression studies with quantitative reverse transcriptase PCR, Western blotting, and lacZ fusions demonstrated that CsrA represses iraD expression. Gel mobility shift, footprint, and toeprint studies identified four CsrA binding sites in the iraD leader transcript, all of which are far upstream of the iraD ribosome binding site. Computational modeling and RNA structure mapping identified an RNA structure that sequesters the iraD Shine-Dalgarno (SD) sequence. Three open reading frames (ORFs), all of which are translated, were identified in the iraD leader region. Two of these ORFs do not affect iraD expression. However, the translation initiation region of the third ORF contains three of the CsrA binding sites, one of which overlaps its SD sequence. Furthermore, the ORF stop codon overlaps the iraD start codon, a sequence arrangement indicative of translational coupling. In vivo expression and in vitro translation studies with wild-type and mutant reporter fusions demonstrated that bound CsrA directly represses translation initiation of this ORF. We further established that CsrA-dependent repression of iraD translation occurs entirely via translational coupling with this ORF, leading to accelerated iraD mRNA decay. IMPORTANCE CsrA posttranscriptionally represses gene expression associated with stationary-phase bacterial growth, often in opposition to the transcriptional effects of the stationary-phase sigma factor RpoS. We show that CsrA employs a novel regulatory mechanism to repress translation of iraD , which encodes an antiadapter protein that protects RpoS against proteolysis. CsrA binds to four sites in the iraD leader transcript but does not directly occlude ribosome binding to the iraD SD sequence. Instead, CsrA represses translation of a short open reading frame encoded upstream of iraD , causing repression of iraD translation via translational coupling. This finding offers a novel mechanism of gene regulation by the global regulator CsrA, and since RpoS can activate csrA transcription, this also highlights a new negative-feedback loop within the complex Csr and RpoS circuitry.

scholarly journals A crustacean Ca2+-binding protein with a glutamate-rich sequence promotes CaCO3 crystallization

2004 ◽  
Vol 384 (1) ◽  
pp. 159-167 ◽  
Author(s):  
Hirotoshi ENDO ◽  
Yasuaki TAKAGI ◽  
Noriaki OZAKI ◽  
Toshihiro KOGURE ◽  
Toshiki WATANABE
Keyword(s):  
Amino Acids ◽  
Binding Sites ◽  
Immunohistochemical Study ◽  
Open Reading Frame ◽  
Reading Frame ◽  
Reverse Transcription Pcr ◽  
Penaeus Japonicus ◽  
Moult Period

The DD4 mRNA of the penaeid prawn Penaeus japonicus was shown previously to be expressed in the epidermis adjacent to the exoskeleton specifically during the post-moult period, when calcification of the exoskeleton took place. The encoded protein possessed a Ca2+-binding site, suggesting its involvement in the calcification of the exoskeleton. In the present study, an additional ORF (open reading frame) of 289 amino acids was identified at the 5′ end of the previous ORF. The newly identified part of the encoded protein included a region of approx. 120 amino acids that was highly rich in glutamate residues, and contained one or more Ca2+-binding sites. In an immunohistochemical study, signals were detected within calcified regions in the endocuticular layer of the exoskeleton. Bacterially expressed partial segments of the protein induced CaCO3 crystallization in vitro. Finally, a reverse transcription-PCR study showed that the expression was limited to an early part of the post-moult period, preceding significant calcification of the exoskeleton. These observations argue for the possibility that the encoded protein, renamed crustocalcin (CCN), promotes formation of CaCO3 crystals in the exoskeleton by inducing nucleation.

ADRENAL FUNCTION IN BREAST CANCER: BIOGENESIS OF ANDROGENS AND CORTISOL BY THE HUMAN ADRENAL GLAND IN VIVO

1970 ◽  
Vol 47 (2) ◽  
pp. 231-242 ◽  
Author(s):  
N. DESHPANDE ◽  
VIBEKE JENSEN ◽  
PAMELA CARSON ◽  
R. D. BULBROOK ◽  
T. W. DOOUSS
Keyword(s):  
Adrenal Gland ◽  
Biosynthetic Pathway ◽  
Adrenal Glands ◽  
Venous Blood ◽  
Minor Amount ◽  
Metabolic Products ◽  
A Minor ◽  
Pool Sizes

SUMMARY A variety of 14C and 3H-labelled steroids have been perfused through the human adrenal gland in situ and their metabolic products isolated from adrenal venous blood. Progesterone, dehydroepiandrosterone and cortisol were isolated after infusion of [3H]pregnenolone; 17α-hydroxyprogesterone, dehydroepiandrosterone and cortisol after infusion of [3H]17α-hydroxypregnenolone and [14C]progesterone; androstenedione and cortisol after infusion of [3H]17α-hydroxyprogesterone and [14C]dehydroepiandrosterone; and 11β-hydroxyandrostenedione after infusion of [3H]androstenedione and [14C]cortisol. From a consideration of the incorporation of radioactivity into the metabolic products, the3H: 14C ratios and the tissue pool sizes it was concluded that the major biosynthetic pathway to cortisol in the human adrenal glands was: pregnenolone→ 17α-hydroxypregnenolone → cortisol. Progesterone was not an important intermediary. Androstenedione was mainly formed by way of 17α-hydroxypregnenolone → 17α-hydroxyprogesterone → androstenedione. 11β-Hydroxyandrostenedione was formed mainly from cortisol and only a minor amount came from the hydroxylation of androstenedione.

scholarly journals In Vitro Identification of Rns-Regulated Genes

2002 ◽  
Vol 184 (4) ◽  
pp. 1196-1199 ◽  
Author(s):  
George P. Munson ◽  
Lisa G. Holcomb ◽  
Heather L. Alexander ◽  
June R. Scott
Keyword(s):  
Escherichia Coli ◽  
Fusion Protein ◽  
Binding Sites ◽  
Genomic Dna ◽  
Maltose Binding Protein ◽  
Open Reading Frame ◽  
Reading Frame ◽  
Fusion Analysis

ABSTRACT To identify Rns-regulated genes, a maltose binding protein (MBP)-Rns fusion protein was used to isolate DNA fragments from enterotoxigenic Escherichia coli genomic DNA that carry Rns binding sites. In vivo transcription fusion analysis shows that Rns positively regulates the expression of the open reading frame of yiiS, which lies immediately downstream of one MBP-Rns-bound fragment.

Electron Probe Analysis of Muscle

1982 ◽  
Vol 40 ◽  
pp. 398-401
Author(s):  
A. V. Somlyo ◽  
H. Shuman ◽  
A. P. Somlyo
Keyword(s):  
Skeletal Muscle ◽  
Smooth Muscle ◽  
Sarcoplasmic Reticulum ◽  
Resting State ◽  
Binding Sites ◽  
Electron Probe ◽  
Frog Skeletal Muscle ◽  
Electron Probe Analysis ◽  
Probe Analysis

Electron probe analysis of frozen dried cryosections of frog skeletal muscle, rabbit vascular smooth muscle and of isolated, hyperpermeab1 e rabbit cardiac myocytes has been used to determine the composition of the cytoplasm and organelles in the resting state as well as during contraction. The concentration of elements within the organelles reflects the permeabilities of the organelle membranes to the cytoplasmic ions as well as binding sites. The measurements of [Ca] in the sarcoplasmic reticulum (SR) and mitochondria at rest and during contraction, have direct bearing on their role as release and/or storage sites for Ca in situ.

Sign in / Sign up

Export Citation Format

Share Document