scholarly journals The insulin enhancer binding site 2 (IEB2; FAR) box of the insulin gene regulatory region binds at least three factors that can be distinguished by their DNA binding characteristics

1995 ◽  
Vol 309 (1) ◽  
pp. 231-236 ◽  
Author(s):  
M L Read ◽  
S B Smith ◽  
K Docherty
Keyword(s):  
Binding Site ◽  
Human Insulin ◽  
Regulatory Region ◽  
Insulin Gene ◽  
Binding Affinities ◽  
Binding Characteristics ◽  
Gene Regulatory ◽  
Equivalent Sequence ◽  
Stimulating Factor

Located at approximately 230 bp upstream from the transcription start site, the insulin enhancer binding site 2 (IEB2) or FAR region of the insulin gene is one of several important sequences involved in regulating transcription of the gene. The present study was undertaken to characterize the transcription factors binding at the IEB2/FAR region of the rat insulin II gene and to compare these with factors known to bind to the equivalent sequence in the rat I and human insulin genes. An endocrine-enriched factor, EFD3, was identified, which bound to the sequence CAGGAG. A second factor (D4) was identified as the widely expressed factor USF (upstream stimulating factor), while a third factor (D5) remained largely uncharacterized. The binding affinities of these three factors differed in the three genes, suggesting that the role of the IEB2/FAR sequence may vary subtly between the rat insulin II, rat insulin I and human insulin genes.

scholarly journals Ligand-specific utilization of the extracellular membrane-proximal region of the gp130-related signalling receptors

1999 ◽  
Vol 345 (1) ◽  
pp. 25-32 ◽  
Author(s):  
Annet HAMMACHER ◽  
John WIJDENES ◽  
Douglas J. HILTON ◽  
Nicos A. NICOLA ◽  
Richard J. SIMPSON ◽  
...  
Keyword(s):  
Proximal Region ◽  
Leukaemia Inhibitory Factor ◽  
Binding Affinities ◽  
Wild Type ◽  
Correct Orientation ◽  
Fibronectin Type Iii ◽  
Membrane Proximal Region ◽  
Stimulating Factor ◽  
Fibronectin Type

The receptor gp130 is used by the interleukin-6 (IL-6)-type cytokines, which include IL-6 and leukaemia-inhibitory factor (LIF). To investigate the role of the three extracellular membrane-proximal fibronectin-type-III-like (FNIII) modules of gp130 and the related receptor for granulocyte colony-stimulating factor (G-CSFR) in cytokine signal transduction we have transfected into murine myeloid M1-UR21 cells the chimaera (GR-FNIII)gp130, which contains the membrane-proximal FNIII modules of the G-CSFR on a gp130 backbone, and its complement, the chimaera (gp130-FNIII)GR. Whereas the binding affinities of 125I-labelled IL-6 to (GR-FNIII)gp130, or of 125I-Tyr1,3-G-CSF to (gp130-FNIII)GR, were similar to wild-type gp130 and wild-type G-CSFR, respectively, 125I-LIF failed to bind with high affinity to (GR-FNIII)gp130. In assays measuring differentiation the (gp130-FNIII)GR cells were fully responsive to G-CSF, whereas the (GR-FNIII)gp130 cells responded fully to the agonistic anti-gp130 monoclonal antibody (mAb) B-S12, but not to IL-6 or LIF. Neutralizing mAbs that recognize the membrane-proximal FNIII modules of gp130 or the G-CSFR differentially interfered with signalling by B-S12, LIF and G-CSF. The data suggest that B-S12 and G-CSF induce the correct orientation or conformation for signalling by the wild-type and chimaeric homodimeric receptors, that the membrane-proximal region of gp130 is important for the correct formation of the signalling IL-6-IL-6 receptor-gp130 complex and that this region is also involved in LIF-dependent receptor heterodimerization and signalling.

scholarly journals The helix-loop-helix transcription factor USF (upstream stimulating factor) binds to a regulatory sequence of the human insulin gene enhancer

1993 ◽  
Vol 295 (1) ◽  
pp. 233-237 ◽  
Author(s):  
M L Read ◽  
A R Clark ◽  
K Docherty
Keyword(s):  
Transcription Factor ◽  
Human Insulin ◽  
Insulin Gene ◽  
Regulatory Sequence ◽  
Mobility Shift ◽  
Helix Loop Helix ◽  
Cell Extracts ◽  
Human Insulin Gene ◽  
Gene Enhancer ◽  
Stimulating Factor

Two important sequence elements, designated insulin enhancer binding site 1 (IEB1) or NIR and IEB2 or FAR, are involved in regulating expression of the rat insulin I gene. These elements bind a helix-loop-helix transcription factor, insulin enhancer factor 1 (IEF1). The IEB1 site is highly conserved among insulin genes but the IEB2 site is not conserved. To investigate the factors binding at the equivalent IEB1 and IEB2 sites in the human insulin gene enhancer, electrophoretic mobility shift assays were performed using a variety of cell extracts and probes specific for the homologous IEB1 and IEB2 sites. The results indicate that a factor with similar tissue distribution and binding characteristics to those of IEF1 binds to the IEB1 site in the human insulin gene, but that a separate factor, identified as the adenovirus major late transcription factor [MLTF, or upstream stimulating factor (USF)] binds to the IEB2 site.

Human insulin gene expression in transgenic mice: mutational analysis of the regulatory region

1996 ◽  
Vol 60 (5) ◽  
pp. 309-316 ◽  
Author(s):  
Jean-Michel Itier ◽  
Philippe Douhet ◽  
Pierrette Desbois ◽  
Rajiv L. Joshi ◽  
Françoise Dandoy-Dron ◽  
...  
Keyword(s):  
Gene Expression ◽  
Transgenic Mice ◽  
Human Insulin ◽  
Mutational Analysis ◽  
Regulatory Region ◽  
Insulin Gene ◽  
Human Insulin Gene ◽  
Insulin Gene Expression

scholarly journals Identification of a Novel PDX-1 Binding Site in the Human Insulin Gene Enhancer

2004 ◽  
Vol 279 (21) ◽  
pp. 22228-22235 ◽  
Author(s):  
John Le Lay ◽  
Taka-aki Matsuoka ◽  
Eva Henderson ◽  
Roland Stein
Keyword(s):  
Binding Site ◽  
Human Insulin ◽  
Insulin Gene ◽  
Human Insulin Gene ◽  
Gene Enhancer

How do histone modifications contribute to transgenerational epigenetic inheritance in C. elegans?

2020 ◽  
Vol 48 (3) ◽  
pp. 1019-1034 ◽  
Author(s):  
Rachel M. Woodhouse ◽  
Alyson Ashe
Keyword(s):  
Histone Modifications ◽  
Molecular Mechanisms ◽  
Epigenetic Inheritance ◽  
Nematode Caenorhabditis Elegans ◽  
Histone Methyltransferases ◽  
Transgenerational Epigenetic Inheritance ◽  
C Elegans ◽  
Recent Developments ◽  
Gene Regulatory

Gene regulatory information can be inherited between generations in a phenomenon termed transgenerational epigenetic inheritance (TEI). While examples of TEI in many animals accumulate, the nematode Caenorhabditis elegans has proven particularly useful in investigating the underlying molecular mechanisms of this phenomenon. In C. elegans and other animals, the modification of histone proteins has emerged as a potential carrier and effector of transgenerational epigenetic information. In this review, we explore the contribution of histone modifications to TEI in C. elegans. We describe the role of repressive histone marks, histone methyltransferases, and associated chromatin factors in heritable gene silencing, and discuss recent developments and unanswered questions in how these factors integrate with other known TEI mechanisms. We also review the transgenerational effects of the manipulation of histone modifications on germline health and longevity.

Lack of association of the polymorphic locus in the 5'-flanking region of the human insulin gene and diabetes in American blacks

Diabetes ◽  
1985 ◽  
Vol 34 (5) ◽  
pp. 433-439 ◽  
Author(s):  
S. Elbein ◽  
P. Rotwein ◽  
M. A. Permutt ◽  
G. I. Bell ◽  
N. Sanz ◽  
...  
Keyword(s):  
Human Insulin ◽  
Polymorphic Locus ◽  
Insulin Gene ◽  
Human Insulin Gene ◽  
Flanking Region ◽  
American Blacks

Cell-specific regulation of IRS-1 gene expression: role of E box and C/EBP binding site in HepG2 cells and CHO cells

Diabetes ◽  
1997 ◽  
Vol 46 (3) ◽  
pp. 354-362 ◽  
Author(s):  
K. Matsuda ◽  
E. Araki ◽  
R. Yoshimura ◽  
K. Tsuruzoe ◽  
N. Furukawa ◽  
...  
Keyword(s):  
Gene Expression ◽  
Binding Site ◽  
Cho Cells ◽  
Hepg2 Cells ◽  
E Box ◽  
Specific Regulation

Cloning and Expression of Recombinant Human Insulin Gene in Pichia pastoris

2019 ◽  
Vol 10 (01) ◽  
Author(s):  
Rafid A. Abdulkareem
Keyword(s):  
Pichia Pastoris ◽  
Human Insulin ◽  
Expression Vector ◽  
Expression System ◽  
Insulin Gene ◽  
Cloning And Expression ◽  
Pcr Analysis ◽  
Major Band ◽  
Human Insulin Gene ◽  
Pichia Pastoris Expression System

The main goal of the current study was cloning and expression of the human insulin gene in Pichia pastoris expression system, using genetic engineering techniques and its treatment application. Total RNA was purified from fresh normal human pancreatic tissue. RNA of good quality was chosen to obtain a first single strand cDNA. Human preproinsulin gene was amplified from cDNA strand, by using two sets of specific primers contain EcoR1 and Notl restriction sites. The amplified preproinsulin gene fragment was double digested with EcoRI and Not 1 restriction enzymes, then inserted into pPIC9K expression vector. The new pPIC9K-hpi constructive expression vector was transformed by the heat-shock method into the E.coli DH5α competent cells. pPic9k –hpi, which was propagated in the positive transformant E. coli cells, was isolated from cells and then linearised by restriction enzyme SalI, then transformed into Pichia pastoris GS115 using electroporation method. Genomic DNA of His+ transformants cell was extracted and used as a template for PCR analysis. The results showed, that the pPic9k – hpi was successfully integrated into the P. pastoris genome, for selected His+ transformants clones on the anticipated band at 330 bp, which is corresponded to the theoretical molecular size of the human insulin gene. To follow the insulin expression in transformans, Tricine–SDS gel electrophoresis and Western blot analysis were conducted. The results showed a successful expression of recombinant protein was detected by the presence of a single major band with about (5.8 KDa) on the gel. These bands correspond well with the size of human insulin with the theoretical molecular weight (5.8 KDa).

Sign in / Sign up

Export Citation Format

Share Document