scholarly journals Characterization of a full-length cDNA encoding human liver S-adenosylmethionine synthetase: tissue-specific gene expression and mRNA levels in hepatopathies

1993 ◽  
Vol 293 (2) ◽  
pp. 481-486 ◽  
Author(s):  
L Alvarez ◽  
F Corrales ◽  
A Martín-Duce ◽  
J M Mato
Keyword(s):  
Amino Acid ◽  
Human Liver ◽  
Structural Features ◽  
Full Length ◽  
Northern Analysis ◽  
Specific Gene ◽  
Mrna Levels ◽  
Calculated Molecular Mass ◽  
Full Length Cdna ◽  
Adenosylmethionine Synthetase

The sequence of a full-length cDNA coding for human liver S-adenosylmethionine synthetase has been determined. It spans 3217 nucleotides and encodes a protein of 395 amino acid residues, with a calculated molecular mass of 43,647 Da. The structural features deduced from the amino acid sequence show a close similarity to those of the rat liver enzyme. The liver-specific S-adenosylmethionine synthetase gene appears to be present as a single copy in the genome, as revealed by Southern analysis. The occurrence of a single mRNA species for this enzyme has been determined by primer extension and Northern analysis. Among several human tissues examined, this gene is expressed only in the liver. Similar S-adenosylmethionine synthetase mRNA levels have been detected in biopsies from normal human liver and from patients with alcoholic cirrhosis and hepatocellular carcinoma. Based on these results, a possible mechanism of regulation of human liver S-adenosylmethionine synthetase is discussed.

scholarly journals Beta-centractin: characterization and distribution of a new member of the centractin family of actin-related proteins.

1994 ◽  
Vol 5 (12) ◽  
pp. 1301-1310 ◽  
Author(s):  
S W Clark ◽  
O Staub ◽  
I B Clark ◽  
E L Holzbaur ◽  
B M Paschal ◽  
...  
Keyword(s):  
Amino Acid ◽  
Cdna Clone ◽  
Expressed Sequence Tag ◽  
Full Length ◽  
Northern Analysis ◽  
Related Protein ◽  
Two Dimensional Gel Electrophoresis ◽  
Full Length Cdna ◽  
Dynactin Complex ◽  
Expressed Sequence

An examination of human-expressed sequence tags indicated the existence of an isoform of centractin, an actin-related protein localized to microtubule-associated structures. Using one of these tags, we isolated and determined the nucleotide sequence of a full-length cDNA clone. The protein encoded represents the first example of multiple isoforms of an actin-related protein in a single organism. Northern analysis using centractin-specific probes revealed three species of mRNA in HeLa cells that could encode centractin isoforms. One mRNA encodes the previously-identified centractin (now referred to as alpha-centractin). The full-length cDNA clone isolated using the expressed sequence tag encodes a new member of the centractin family, beta-centractin. A probe specific for alpha-centractin hybridized to the third species of mRNA observed (referred to as gamma-centractin). Comparisons of Northern blots of human tissues indicated that alpha-centractin and beta-centractin mRNAs are equally distributed in all populations of mRNA examined, whereas the expression of gamma-centractin appears to be tissue specific. The amino acid sequence of beta-centractin, deduced from the cDNA, indicates a 91% identity with alpha-centractin, increasing to 96% similarity when conservative amino acid changes are taken into account. As antibodies previously raised against alpha-centractin reacted only poorly with beta-centractin, new antibodies were produced and combined with two-dimensional gel electrophoresis to discriminate the two isoforms. Using this system, the subcellular distribution of the alpha- and beta-isoforms were determined. Both isoforms were found predominantly in the cytosolic fraction as a part of a previously identified 20S complex (referred to as the dynactin complex) with no evidence for a free pool of either isoform. The isoforms were found in a constant ratio of approximately 15:1 (alpha:beta) in the dynactin complex.

Molecular Cloning, Characterisation, Tissue Distribution, and mRNA Expression Changes under Different Light Regimes of Brain and Muscle Arnt-Like Protein-1 (BMAL1) in Columba Livia

2017 ◽  
Vol 10 (3) ◽  
pp. 156-163 ◽  
Author(s):  
Y. Wang ◽  
H.M. Yang ◽  
Y.B. Li ◽  
W. Cao
Keyword(s):  
Circadian Rhythm ◽  
Light Exposure ◽  
Columba Livia ◽  
Full Length ◽  
Day Length ◽  
Mrna Levels ◽  
Tyto Alba ◽  
Full Length Cdna ◽  
Light Regimes ◽  
Significant Difference

Brain and muscle Arnt-like protein-1 (BMAL1) plays an important role in circadian rhythm, which is involved in daily behaviours and physiological activities. However, little is known about the molecular function of BMAL1 in the Pigeon ( Columba livia). In our study, the full-length cDNA of Bmal1 was cloned and sequenced from the Pigeon for the first time, and submitted to the GenBank to obtain the accession number (KF906247). The full-length cDNA of Bmal1 consists of 2,488 nucleotides, and encodes 634 amino acids. Phylogenetic analysis showed that it bore the greatest similarity to Bmal1 from the Chicken ( Gallus gallus) and Barn Owl ( Tyto alba). The amino acid sequence of the Pigeon BMAL1 contained a HLH domain and two PAS domains, which are involved in forming hetero-homodimers with the CLOCK as the positive element of the circadian rhythm. The results of real-time quantitative PCR of Bmal1 under different light regimes showed that the amplitude and expression pattern of Bmal1 were strongly affected by day length. Bmal1 was most highly expressed in the pancreas. Relative to Bmal1 expression level under 12 h of light exposure, it was increased significantly in the pituitary gland, ovary and uterus under 15 h of light exposure ( P < 0.05). However, other tissues, including the hypothalamus, heart, liver, spleen, kidney, intestines, crureus, and pectorals exhibited no significant difference ( P < 0.05) under the two light regimes. This is the first study to investigate Bmal1 mRNA levels in various tissues under different light cycles, and thereby provide data for further study of the molecular and regulatory mechanisms of Bmal1 and circadian clock genes of the Pigeon.

Cloning, characterisation and expression profile of kisspeptin1 and the kisspeptin1 receptor in the hypothalamic–pituitary–ovarian axis of Chinese alligator Alligator sinensis during the reproductive cycle

2020 ◽  
Vol 32 (8) ◽  
pp. 792 ◽  
Author(s):  
Ruidong Zhang ◽  
Haitao Nie ◽  
Shulong Duan ◽  
Peng Yan ◽  
Ali Izaz ◽  
...  
Keyword(s):  
Amino Acid ◽  
Quantitative Polymerase Chain Reaction ◽  
Reproductive Period ◽  
Full Length ◽  
Reading Frame ◽  
Full Length Cdna ◽  
Cdna Sequences ◽  
Acid Protein ◽  
Amino Acid Precursor ◽  
Alligator Sinensis

Kisspeptin1 (Kiss1), a product of the Kiss1 gene, plays an important role in the regulation of reproduction in vertebrates by activating the Kiss1 receptor (Kiss1R) and its coexpression with gonadotrophin-releasing hormone (GnRH) in GnRH neurons. The purpose of this study was to clone the Kiss1 and Kiss1R genes found in the brain of Alligator sinensis and to explore their relationship with reproduction. The full-length cDNA of Kiss1 is 816bp, the open reading frame (ORF) is 417bp and the gene encodes a 138-amino acid precursor protein. The full-length cDNA of Kiss1R is 2348bp, the ORF is 1086bp and the gene encodes a 361-amino acid protein. Quantitative polymerase chain reaction showed that, except for Kiss1R expression in the hypothalamus, the expression of Kiss1 and Kiss1Rduring the reproductive period of A. sinensis was higher than that in the hypothalamus, pituitary gland and ovary during the hibernation period. The changes in GnRH2 mRNA in the hypothalamus were similar to those of GnRH1 and peaked during the reproductive period. This study confirms the existence of Kiss1 and Kiss1R in A. sinensis and the findings strongly suggest that Kiss1 and Kiss1R may participate in the regulation of GnRH secretion in the hypothalamus of alligators during the reproductive period. Furthermore, this is the first report of the full-length cDNA sequences of Kiss1 and Kiss1R in reptiles.

A Novel Human Actin-Binding Protein Homologue That Binds to Platelet Glycoprotein Ib

Blood ◽  
1998 ◽  
Vol 92 (4) ◽  
pp. 1268-1276 ◽  
Author(s):  
Wen-feng Xu ◽  
Zhi-wei Xie ◽  
Dominic W. Chung ◽  
Earl W. Davie
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Binding Protein ◽  
Full Length ◽  
Actin Binding ◽  
Intracellular Domain ◽  
Platelet Surface ◽  
Full Length Cdna ◽  
Actin Binding Protein ◽  
Amino Terminal

Glycoprotein (GP)Ib-IX-V is one of the major transmembrane complexes present on the platelet surface. Its extracellular domain binds von Willebrand factor (vWF) and thrombin, while its intracellular domain associates tightly with the cytoskeleton through the actin-binding protein (ABP)-280, also known as filamin. In the present study, a full-length cDNA coding for a human ABP homologue has been cloned and sequenced. This protein was identified by the yeast two-hybrid screening procedure via its interaction with the intracellular domain of GPIb. Initially, a 1.3-kb partial cDNA was isolated from a megakaryocyte-like cell line (K562) cDNA library followed by a full-length cDNA of 9.4 kb that was identified in a human placenta library. The full-length cDNA encoded a protein of 2,578 amino acids with a calculated molecular weight of 276 kD (ABP-276). The amino terminal 248 amino acids contained an apparent actin binding domain followed by 24 tandem repeats each containing about 96 amino acids. The amino acid sequence of the protein shared a high degree of homology with human endothelial ABP-280 (70% identity) and chicken filamin (83% identity). However, the 32 amino acid Hinge I region in ABP-280 that contains a calpain cleavage site conferring flexibility on the molecule, was absent in the homologue. An isoform containing a 24 amino acid insertion with a unique sequence at the missing Hinge I region was also identified (ABP-278). This isoform resulted from alternative RNA splicing. ABP-276 and/or ABP-278 were present in all tissues examined, but the relative amount varied in that some tissue contained both forms, while other tissue contained predominately one or the other. © 1998 by The American Society of Hematology.

scholarly journals Isolation, characterization and sequence analysis of a full-length cDNA clone encoding acetohydroxy acid reductoisomerase from spinach chloroplasts

1991 ◽  
Vol 277 (2) ◽  
pp. 469-475 ◽  
Author(s):  
R Dumas ◽  
M Lebrun ◽  
R Douce
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Single Gene ◽  
Amino Acid Sequences ◽  
Full Length ◽  
Open Reading Frame ◽  
Amino Acid Residues ◽  
Protein Precursor ◽  
Reading Frame ◽  
Full Length Cdna

Acetohydroxy acid reductoisomerase (AHRI), the second enzyme in the parallel isoleucine/valine-biosynthetic pathway, catalyses an unusual two-step reaction in which the substrate, either 2-acetolactate or 2-aceto-2-hydroxybutyrate, is converted via an alkyl migration and an NADPH-dependent reduction to give 2,3-dihydroxy-3-methylbutyrate or 2,3-dihydroxy-3-methylvalerate respectively. We have isolated and characterized a full-length cDNA from a lambda gt11 spinach library encoding the complete acetohydroxy acid reductoisomerase protein precursor. The 2050-nucleotide sequence contains a 1785-nucleotide open reading frame. The derived amino acid sequence indicates that the protein precursor consists of 595 amino acid residues including a presequence peptide of 72 amino acid residues. The N-terminal sequence of the first 16 amino acid residues of the purified AHRI confirms the identity of the cDNA. The derived amino acid sequence from this open reading frame shows 23% identity with the deduced amino acid sequences of the Escherichia coli and Saccharomyces cerevisiae AHRI proteins. There are two blocks of conserved amino acid residues in these three proteins. One of these is a sequence similar to the ‘fingerprint’ region of the NAD(P)H-binding site found in a large number of NAD(P)H-dependent oxidoreductases. The other, a short sequence (Lys-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Ser-His-Gly-Phe) containing the amino acids lysine and histidine, could well be the catalytic site of the first step of the AHRI reaction. Southern-blot analysis indicated that AHRI is encoded by a single gene per haploid genome of about 7.5 kbp containing at least four introns.

scholarly journals Structure of a full-length cDNA clone for the preproα2(I) chain of human type I procollagen. Comparison with the chicken gene confirms unusual patterns of gene conservation

1988 ◽  
Vol 252 (3) ◽  
pp. 633-640 ◽  
Author(s):  
H Kuivaniemi ◽  
G Tromp ◽  
M L Chu ◽  
D J Prockop
Keyword(s):  
Amino Acid ◽  
Cdna Clone ◽  
Full Length ◽  
Total Amino Acid ◽  
Type I ◽  
Amino Acid Residues ◽  
Full Length Cdna ◽  
Human Type ◽  
Type I Procollagen ◽  
I Gene

A cDNA clone from a human placental library was found to consist of an essentially full-length cDNA of 4.6 kb for the prepro alpha 2(I) chain of type I procollagen. Nucleotide sequencing of the 5′-end of the cDNA provided a sequence of 1617 nucleotide residues and codons for 539 amino acid residues not previously defined. Comparison of the complete structure of the prepro alpha 2(I) cDNA with previously reported sequences for the chicken pro alpha 2(I) gene indicated that 83% of 1366 total amino acid residues were conserved. In the alpha-chain domain 84% of 1014 amino acid residues were conserved. Also, there was conservation of the previously noted preference for U and C in the third position of codons for glycine, proline and alanine. One major difference between the human and the chicken prepro alpha 2(I) chain was that the human chain contained 21 fewer proline residues, an observation that probably explains why the triple helix of human type I procollagen unfolds at temperatures that are 1-2 degrees C lower. In parallel experiments, sequencing of intron-exon boundaries for nine exons of genomic subclones confirmed and extended previous observations that the pro alpha 2(I) gene, like other genes from fibrillar collagens, has an unusual 54-base pattern of exon sizes that is highly conserved through evolution.

scholarly journals The complete amino acid sequence of a pea (Pisum sativum) seed lipoxygenase predicted from a near full-length cDNA

1988 ◽  
Vol 253 (3) ◽  
pp. 915-918 ◽  
Author(s):  
P M Ealing ◽  
R Casey
Keyword(s):  
Amino Acid ◽  
Pisum Sativum ◽  
Amino Acid Sequence ◽  
Amino Acid Identity ◽  
Full Length ◽  
Predicted Amino Acid Sequence ◽  
Protein Coding ◽  
Full Length Cdna ◽  
Coding Regions

A near full-length cDNA for a pea (Pisum sativum) seed lipoxygenase was isolated and sequenced. It has a protein coding sequence (2583 bp), 5′ (59 bp) and 3′ (191 bp) non-coding regions, and a poly(A) tail (20 bp). The predicted amino acid sequence indicates a polypeptide of Mr 97,628 that shows about 86% amino acid identity with a soya-bean lipoxygenase 3 protein sequence [Yenofsky, Fine & Liu (1988) Mol. Gen. Genet. 211, 215-222]. The cDNA directs the transcription of mRNA that can be translated to give an anti-lipoxygenase-precipitable polypeptide in vitro.

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