Molecular cloning and expression of rat antisecretory factor and its intracellular localization

1999 ◽  
Vol 77 (3) ◽  
pp. 223-228 ◽  
Author(s):  
Kayoko Tateishi ◽  
Yoshio Misumi ◽  
Yukio Ikehara ◽  
Kyoko Miyasaka ◽  
Akihiro Funakoshi
Keyword(s):  
Pituitary Gland ◽  
Intestinal Mucosa ◽  
Intracellular Localization ◽  
Cloning And Expression ◽  
Cdna Libraries ◽  
Cdna Clones ◽  
Cytoplasmic Protein ◽  
Rat Pituitary ◽  
26S Protease ◽  
Antisecretory Factor

Antisecretory factor (AF) was identified as a pituitary protein that inhibits the intestinal fluid secretion induced by cholera toxin. One aim of this study was to elucidate whether AF is also synthesized in the intestine or if AF produced in the pituitary is transported to the intestinal tract for its function there. cDNA clones encoding a protein proposed to be AF were isolated from rat pituitary gland and intestinal mucosa cDNA libraries. The nucleotide sequences of clones isolated from the rat pituitary gland and intestinal mucosa were identical. The deduced amino acid sequence was highly homologous to the sequence for subunit 5a of the human 26S protease that exists abundantly in the cytosol and nucleus. The production of AF in the intestine was confirmed by Northern blot and immunoblot analyses. Immunocytochemical observations of cells transfected with the rat AF cDNA showed that the AF protein was localized in the cytoplasm. These findings suggest that the protein proposed to be AF may be a cytoplasmic protein, it exists in the intestine rather than being transported from the pituitary gland, and it may function in intestinal cells.Key words: rat antisecretory factor, 26S protease, S5a, cytoplasmic protein.

scholarly journals Molecular cloning and expression of the cDNA for alpha 3 subunit of human alpha 3 beta 1 (VLA-3), an integrin receptor for fibronectin, laminin, and collagen.

1991 ◽  
Vol 115 (1) ◽  
pp. 257-266 ◽  
Author(s):  
Y Takada ◽  
E Murphy ◽  
P Pil ◽  
C Chen ◽  
M H Ginsberg ◽  
...  
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Transmembrane Domain ◽  
Chinese Hamster Ovary Cells ◽  
General Structure ◽  
Chinese Hamster ◽  
Cloning And Expression ◽  
Cdna Libraries ◽  
Cdna Clones ◽  
Alpha Subunits

alpha 3 beta 1 (VLA-3), a member of the integrin family of cell adhesion receptors, may function as a receptor for fibronectin, laminin, and collagen. A partial cDNA clone (2.4 kb) for the human alpha 3 subunit was selected from an endothelial cell lambda gt11 cDNA library by specific antibody screening. Several overlapping cDNA clones were subsequently obtained, of a total length of 4.6 kb from various cDNA libraries. The reconstructed alpha 3 cDNA was expressed on the surface of chinese hamster ovary cells as detected by an alpha 3-specific mAb after transfection, suggesting that the cDNA is authentic. Within this sequence was an open reading frame, encoding for 1,051 amino acids, including a signal peptide of 32 residues, a long extracellular domain (959 residues), a transmembrane domain (28 residues), and a short cytoplasmic segment (32 residues). Overall, the alpha 3 amino acid sequence was 25-37% similar to the other integrin alpha subunits that are cleaved, with most similarity to the alpha 6 sequence (37%), and less similarity to those alpha subunits that have I domains (15-20%, excluding the I domain sequence itself). Features most like those in other alpha subunits are (a) the positions of 18/19 cysteine residues, (b) three potential metal binding domains of the general structure DX(D/N)X(D/N)GXXD, and (c) the predicted transmembrane domain. The mass of alpha 3 calculated from its amino acid sequence is 113,505. The human alpha 3 sequence was 89% identical to hamster galactoprotein b3, and 70% similar to the chicken CSAT antigen band 2 protein partial sequence, suggesting that these two polypeptides are homologues of human alpha 3.

scholarly journals CircAgtpbp1 Acts as a Molecular Sponge of miR-543-5p to Regulate the Secretion of GH in Rat Pituitary Cells

Animals ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 558
Author(s):  
ZeWen Yu ◽  
WenZhi Ren ◽  
Tian Wang ◽  
WeiDi Zhang ◽  
ChangJiang Wang ◽  
...  
Keyword(s):  
Pituitary Gland ◽  
Inhibitory Effect ◽  
Pituitary Cells ◽  
Sequencing Analysis ◽  
Hormone Regulation ◽  
Gh Secretion ◽  
Qrt Pcr ◽  
Rat Pituitary ◽  
Rat Pituitary Cells

CircRNAs have been identified to be expressed differently and stably in numerous species and tissues, but their functions in growth hormone (GH) secretion are still largely unknown. In summary, we have revealed a circRNA-miRNA-mRNA network that may play a biological role in the rat pituitary gland. First, we verified the chromosome location information of circAgtpbp1 according to sequencing analysis. The circAgtpbp1 characteristics were authenticated through PCR, qRT–PCR, treating with RNase and fluorescent in situ hybridization (FISH). Second, we detected the expression pattern of circAgtpbp1 in the rat anterior pituitary by qRT–PCR. We also designed circAgtpbp1 siRNA and constructed overexpression plasmid to evaluate the effect of circAgtpbp1 function on GH secretion by qRT–PCR, ELISA and Western blot. CircAgtpbp1 is a stable, truly circular molecule. We found that circAgtpbp1 interacted with miR-543-5p and can regulate GH secretion in pituitary cells through a circAgtpbp1-miR-543-5p-GH axis. Overall, the evidence generated by our study suggests that circAgtpbp1 can act as a sponge of miR-543-5p to reduce the inhibitory effect of miR-543-5p on Gh1 and further promote GH secretion. These findings expand our existing knowledge on the mechanisms of hormone regulation in the pituitary gland.

Synaptophysin Immunoreactivity in the Aging Rat Pituitary Gland

1997 ◽  
Vol 43 (6) ◽  
pp. 561-564 ◽  
Author(s):  
L.C Saland ◽  
A Apodaca ◽  
D Ramirez ◽  
V Hernandez ◽  
J Gaddy ◽  
...  
Keyword(s):  
Pituitary Gland ◽  
Rat Pituitary ◽  
Aging Rat

DISTRIBUTION OF NEUROSECRETORY GRANULES AMONG THE ANATOMICAL COMPARTMENTS OF THE NEUROSECRETORY PROCESSES OF THE PITUITARY GLAND: A QUANTITATIVE ULTRASTRUCTURAL APPROACH TO HORMONE STORAGE IN THE NEURAL LOBE

1976 ◽  
Vol 68 (2) ◽  
pp. 225-NP ◽  
Author(s):  
J. F. MORRIS
Keyword(s):  
Basement Membrane ◽  
Pituitary Gland ◽  
Neurosecretory Granules ◽  
Neural Lobe ◽  
Releasable Pool ◽  
Readily Releasable Pool ◽  
Rat Pituitary ◽  
Axonal Swellings ◽  
Granule Release

SUMMARY The distribution of neurosecretory granules in various anatomical compartments of neurosecretory axons of the neural lobe of the rat pituitary has been studied. Apart from the most anterior tip of the gland, where granules are largely restricted to undilated axons and a few 'swellings', the proportional compartmental storage of granules is essentially homogeneous for the rest of the gland: 13% of granules are found in undilated axons, 31% in axonal 'endings' (which contain microvesicles and abut the basement membrane) and 55% in axonal 'swellings' (which are devoid of significant numbers of microvesicles). These values indicate that the 'endings' contain a much greater proportion of the total number of granules stored in the neural lobe than would be predicted if the readily releasable pool of hormone were composed of all the granules in the 'endings'. Some further constraint on granule release either physiological or anatomical (e.g. the position of the granule in relation to the plasmalemma of the 'ending') must be operative.

scholarly journals Cloning and sequence analysis of cDNAs encoding the α1, α2 and α3 chains of mouse collagen VI

1993 ◽  
Vol 291 (3) ◽  
pp. 787-792 ◽  
Author(s):  
R Z Zhang ◽  
T C Pan ◽  
R Timpl ◽  
M L Chu
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequences ◽  
Untranslated Regions ◽  
Cdna Libraries ◽  
Cdna Clones ◽  
Collagen Vi ◽  
Central Segment ◽  
Glycosylation Sites ◽  
Key Features ◽  
Triple Helical Domain

cDNA clones encoding the alpha 1, alpha 2 and alpha 3 chains of mouse collagen VI have been isolated by screening cDNA libraries with the corresponding human probes. The composite cDNAs for the alpha 1, alpha 2, and alpha 3 chains are 2.5, 1.6 and 2.9 kb in size respectively. The alpha 1 and alpha 2 cDNAs encode the C-terminal portions of the chains as well as the entire 3′-untranslated regions, while the alpha 3 cDNAs encode a central segment of 959 amino acids flanking the triple-helical domain. The deduced amino acid sequences share 86-88% identity with the human counterparts and 67-73% identity with the chicken equivalents. Alignment of the deduced amino acid sequences of mouse, human and chicken collagens reveal that the key features of the protein, including the cysteine residues, imperfections in the Gly-Xaa-Xaa regions, Arg-Gly-Asp sequences and potential N-glycosylation sites, are mostly conserved.

scholarly journals A new intracellular serine protease inhibitor expressed in the rat pituitary gland complexes with granzyme B

FEBS Letters ◽  
1998 ◽  
Vol 440 (3) ◽  
pp. 361-364 ◽  
Author(s):  
Rena M Hill ◽  
Kate S Morresey ◽  
Leigh C Coates ◽  
Eva Mezey ◽  
Brennan Fell ◽  
...  
Keyword(s):  
Protease Inhibitor ◽  
Pituitary Gland ◽  
Serine Protease ◽  
Granzyme B ◽  
Serine Protease Inhibitor ◽  
Rat Pituitary

An RNA-binding protein from Xenopus oocytes is associated with specific message sequences

Development ◽  
1987 ◽  
Vol 101 (4) ◽  
pp. 741-749 ◽  
Author(s):  
D.R. Crawford ◽  
J.D. Richter
Keyword(s):  
Xenopus Oocytes ◽  
Sucrose Gradient ◽  
Rna Binding ◽  
Binding Protein ◽  
Rna Binding Protein ◽  
Cdna Libraries ◽  
Cdna Clones ◽  
Stage Of Development ◽  
Messenger Ribonucleoprotein ◽  
Stage 1

Monoclonal antibodies directed against an RNA-binding protein from Xenopus oocytes were used to immunoselect messenger ribonucleoprotein (mRNP) particles. RNA was extracted from both the immunoselected and nonselected fractions and was used to direct the synthesis of oligo (dT)-primed 32P-cDNA. These two cDNA preparations were then used to probe Xenopus stage-1 oocyte cDNA libraries to identify sequences that had been specifically coimmunoselected by the antibodies. Three cDNA clones were shown to be derived specifically from the antibody-selected mRNPs. During very early oogenesis (stage 1–2), the RNA-binding protein and the three coselected mRNAs sediment in the nontranslating mRNP region of a sucrose gradient. By oocyte stage 6, the binding protein concentration decreases by as much as 22-fold relative to polyadenylated RNA. At this stage of development, the three mRNAs are found predominantly in the polysome region of a sucrose gradient. These data demonstrate that Xenopus oocytes contain an RNA-binding protein which binds specific message sequences and may regulate their expression.

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