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1) Hormones like steroids and thyroid hormones act via nuclear receptors, which are ligand-activated transcription factors that reside in the cell nucleus.
2) When a hormone binds its receptor, it causes the receptor to form a dimer that then binds to DNA at hormone response elements and regulates gene transcription.
3) In addition to direct DNA binding, nuclear receptors can also regulate transcription through protein-protein interactions with other transcription factors.
1) Hormones like steroids and thyroid hormones act via nuclear receptors, which are ligand-activated transcription factors that reside in the cell nucleus.
2) When a hormone binds its receptor, it causes the receptor to form a dimer that then binds to DNA at hormone response elements and regulates gene transcription.
3) In addition to direct DNA binding, nuclear receptors can also regulate transcription through protein-protein interactions with other transcription factors.
1) Hormones like steroids and thyroid hormones act via nuclear receptors, which are ligand-activated transcription factors that reside in the cell nucleus.
2) When a hormone binds its receptor, it causes the receptor to form a dimer that then binds to DNA at hormone response elements and regulates gene transcription.
3) In addition to direct DNA binding, nuclear receptors can also regulate transcription through protein-protein interactions with other transcription factors.
receptors, some (mostly hormones) have intracellular receptors. The ligands of such receptors are small hydrophobic molecules, and binding of the appropriate ligand converts them into active transcription factors, namely, proteins which stimulate, or reduce, the rate of gene transcription. HORMONES WITH NUCLEAR RECEPTORS (continued)
Thus these receptors, collectively
named nuclear receptors (NRs), function as ligand-activated transcription factors. The five types of steroid hormones have nuclear receptors, and so do the thyroid hormones. In addition, derivatives of vitamin D and vitamin A (retinoids), which are synthesized from precursors consumed as food, also have NRs. STEROID VERSUS NON-STEROID LIGANDS
The ligands shown in the next slide are
divided into subgroups: steroids (left column) and non-steroids (middle column). The right column shows the names of the receptors of the compounds presented. Some receptors are named after a group of compounds (natural or synthetic), all of which can activate that receptor (e.g., estrogen receptor), rather than after a specific compound. HORMONES WITH NUCLEAR RECEPTORS 11-CIS- RETINAL IN THE RETINA: ACTION NOT MEDIATED BY A RETINOID RECEPTOR MECHANISM OF RECEPTOR ACTIVATION
The inactive receptors of four of the
steroids: cortisol, aldosterone, progesterone and testosterone (GR, MR, PR and AR) reside in the cytoplasm. A single receptor molecule forms a complex with several proteins, most of them heat- shock proteins (HSPs). The composition of the complexes of the various receptors vary somewhat, but all of them include HSP90. HSPs are known to be induced by various cellular stresses, and to act as chaperons, namely, proteins that act to preserve the conformation of other proteins. RECEPTOR ACTIVATION (Continued)
When one of the four steroid hormones
binds to its receptor, the complex disintegrates; two activated receptor molecules form a homodimer, which enters the nucleus and functions as a transcription factor. The inactive estradiol receptor resides in the nucleus as a dimer, in a complex with one HSP90 molecule, which dissociates upon binding of the hormone. ACTIVATION OF STEROID HORMONE RECEPTORS RECEPTOR ACTIVATION (Continued)
The inactive receptors of thyroid
hormones, vitamin D and the retinoids reside in the nucleus. Upon hormone binding they form dimers. TR regulates a few genes as a homodimer, but in most cases it acts as a heterodimer with RXR. VDR and RAR act primarily as heterodimers with RXR. In some cases the inactive receptor binds to promoters and act in an opposite manner to that of the activated receptor (inhibition instead of activation). STRUCTURE OF NUCLEAR RECEPTORS (NRs) (see explanations below) STRUCTURE OF NUCLEAR RECEPTORS
Nuclear receptors are homologous to a
variable extent. It is customary to divide the receptor molecule into five domains: A/B, C, D, E, and F. Domain C, the DNA-binding domain, shows the highest degree of homology among the receptors. Domain E, the ligand-binding domain, shows lower degree of homology. Greater homology is observed when the ligands are similar. STRUCTURE OF NUCLEAR RECEPTORS (Continued)
The transactivation domains, namely,
those involved in affecting the transcription machinery, are A/B and E (E is ligand-dependent). They are named AF-1 and AF-2, respectively. (AF=activating function). The above figure shows also the sequences responsible for other functions of the receptor: dimerization, HSP binding, and nuclear localization. THE TWO ZINC FINGERS IN THE DNA- BINDING DOMAIN (DBD) STRUCTURE OF A HOMODIMER OF THE DNA-BINDING DOMAIN OF A NUCLEAR RECEPTOR: THE PROXIMAL (N- TERMINAL) ZINC FINGERS OF THE MONOMERS FIT CONSECUTIVE TURNS IN THE MAJOR GROOVE OF THE DNA DOUBLE HELIX. THE DISTAL ZINC FINGERS (INCLUDED IN THE DIMERIZATION ZONE) LIE OVER THE MINOR GROOVE. REGULATION OF TRANSCRIPTION REGULATION OF TRANSCRIPTION (PRINCIPLES)
The above figure illustrates the
regulation of transcription in animal cells. Sections of the plasma membrane, and the outline of the nuclear membrane, are shown schematically. RNA polymerase II (Pol II), which executes transcription, is shown as part of a complex with other proteins (general transcription factors). REGULATION OF TRANSCRIPTION (Continued)
This complex, the basal transcription
complex, is bound to a sequence of bases in the DNA, upstream of the transcription initiation site. In most genes this DNA site includes the sequence TATA, and is named the TATA box. The basal transcription complex interacts with various proteins which bind to the DNA, usually upstream of the polymerase complex. These proteins (transcription factors - TFs) regulate (increase or decrease) the rate of transcription. REGULATION OF TRANSCRIPTION (Continued)
The promoter of a gene is the DNA
section upstream of the transcription initiation site, and includes both the RNA polymerase complex binding site and the DNA section which binds the transcription factors (a couple of thousands basepairs may be relevant). Transcription factors bind selectively to certain DNA sequences (response elements REs). REGULATION OF TRANSCRIPTION (Continued)
The above figure shows examples of
two types of TFs: ligand-activated (a steroid hormone receptor), and phosphorylation-activated (substrates of protein kinases, which had been activated as a consequence of increases in second messengers). Certain TFs are known to be activated by dephosphorylation rather than by phosphorylation. REGULATION OF TRANSCRIPTION (Continued)
Usually, transcription factors affect the
Pol II complex indirectly, with the mediation of proteins named coactivators or corepressors, depending on the effect on transcription. The figure shows a mechanism in which the steroid hormone receptors need two coactivators in a row. The second coactivator (CBP) mediates also the action of other TFs. REGULATION OF TRANSCRIPTION (Continued)
Coactivators are activated allosterically
by TFs and in turn affect allosterically proteins down the chain. Some coactivators (in the figure: CBP) also stimulate transcription by another mechanism: they have enzymatic activity of histone acetyl transferase (HAT). Acetylation of certain lysine residues in histones loosens the tight chromatin structure, thus facilitating transcription. REGULATION OF TRANSCRIPTION (Continued)
Similarly, corepressors exert allosteric
activation on histone deacetylases (HDACs), which have an opposite effect on transcription. RESPONSE ELEMENTS FOR THE STEROID HORMONE RECEPTORS
ACTGGA PALINDROME (INVERTED REPEAT) AGGTCA TGACCT
DIRECT REPEAT AGGTCA AGGTCA RESPONSE ELEMENTS OF NRs
As shown in the above figure, dimers of
NRs bind to response elements comprised of two half-sites; each half-site binds one monomer. One type of response element is a palindrome (inverted repeat), in which the half-sites, one a mirror image of the other regarding the sequence of bases, are found in each DNA strand. This is the most common response element for steroid hormone receptor homodimers. RESPONSE ELEMENTS OF NRs (continued)
The other type of response element is a
direct repeat, in which the same base sequence appears twice in a row on the same DNA strand. The specificity of a response element for a receptor is determined not only by the base sequence but also by the spacing between the two half-sites. It should be stressed that TFs other than NRs are often homo- or heterodimers, and their response elements are also of the above types. ADDITIONAL MODES OF ACTION OF TRANSCRIPTION FACTORS
As described above, many TFs act via direct
binding to the DNA. However, an alternative mode of action is binding by protein-protein interaction to another TF, which in turn is bound to the DNA. In such case, the TF which does not bind to DNA directly may either augment or suppress the action of the DNA-bound TF. Another mode of action is the binding of a TF to another TF, thus excluding the association of the latter with the DNA. ADDITIONAL MODES OF ACTION OF TRANSCRIPTION FACTORS (Continued)
Some TFs can act via two of the described
mechanisms, or even by all three of them. This pertains to some nuclear receptors. Most of the actions of the glucocorticoid receptor (GR) to suppress the immune system involve protein-protein interaction with other TFs (second and third mechanism). The estrogen and androgen receptors were also shown to bind to DNA indirectly (via other TFs) in some promoters. ADDITIONAL MODES OF ACTION OF TRANSCRIPTION FACTORS (Continued)
When the activated receptors of steroid
hormones act on transcription via protein-protein interaction, they act as monomers rather than as dimers. ORPHAN RECEPTORS
After several nuclear receptors had
been cloned, investigators looked for additional homologous proteins at the RNA level. Most of the novel NRs found had no known ligands, and thus were named orphan receptors. Eventually, ligands were identified for some orphan receptors. These ligands included regulatory molecules arriving from the outside, as well as metabolites synthesized within the cell. ORPHAN RECEPTORS (Continued)
The ligands identified for orphan receptors
are hydrophobic, as are the ligands of the classical NRs. Some of the orphan receptors may be regulated by phosphorylation rather than by ligands. There is great interest in the orphan receptors, especially since some drugs used in clinical medicine, as well as some pollutants, act via orphan receptors.
(Dermatology-Laboratory and Clinical Research - Cell Biology Research Progress) Xiao-Peng Ma, Xiao-Xiao Sun - Melanin - Biosynthesis, Functions and Health Effects-Nova Science Publishers (2012)