Post translational time stamping is present from cyanobacteria to mammals. All organisms have evolved timing mechanisms to adapt to environmental changes in order to optimize survival and improve fitness for an environment. To anticipate these regular daily electromagnetic cycles of light and dark, many organisms manifest near 24-h cell-autonomous oscillations that are sustained by transcription–translation-based or post-transcriptional negative feedback loops that control a wide range of biological processes. With an eye to identifying emerging common themes among cyanobacterial, fungal and animal clocks, some major recent developments in the understanding of the mechanisms that regulate these oscillators and their output need to be discussed. These include roles for antisense transcription, intrinsically disordered proteins, codon bias in clock genes, and a more focused discussion of post-transcriptional and translational regulation as a part of both the oscillator and output.
Circadian rhythms in every organism are cell autonomous, appear to have arisen only a few times in evolution, and can be driven by one of a few lineage-specific but otherwise highly conserved central oscillators. While oscillators driving bacterial and plant clocks are distinct from each other and from other known clocks, fungal and animal cells share circadian oscillators of conserved regulatory architecture: transcription-translation feedback loops (TTFLs) comprised of two parts.
Specifically, 1) a positive arm with a heterodimeric complex at its core that behaves as the activator of the system, promoting the transcription of 2) one or more components of the negative arm, which when translated inhibit the activity of the positive arm.
WHAT IS THE TTFL?
Transcription-translation feedback loop (TTFL) is THE cellular model for explaining circadian rhythms in behavior and physiology. It is widely conserved across species, and the TTFL is largely auto-regulatory with the assistance of the sun & moon and dark periods on Earth, in which transcription of clock genes is regulated by their own protein products. This implies that light and dark control genetic expression and not the other way around. The TTFL is a negative feedback loop, in which clock genes are regulated by their protein products. Generally, the TTFL involves 2 main arms: positive regulatory elements that promote transcription and protein products that suppress transcription. When a positive regulatory element binds to a clock gene promoter, transcription of DNA proceeds, resulting in the creation of an mRNA transcript, and then translation proceeds, resulting in a semiconductive protein product. There are characteristic delays between mRNA transcript accumulation, protein accumulation, and gene suppression due to translation dynamics, post-translational protein modification, protein dimerization, and intracellular travel to the nucleus. Across species, proteins involved in the TTFL contain common structural motifs such PAS domains, involved in protein-protein interactions, and bHLH domains, involved in DNA binding.

The two overarching areas characteristic of circadian systems in general: 1) the negative arm and its regulation of the core clock; 2) the control of output by the positive arm and its environment.
In ALL mammals the heterodimeric BMAL1-CLOCK complex positively regulates expression of negative arm component genes, the Periods and Cryptochromes(encoding PER1, PER2, PER3, CRY1 and CRY2), that combine with CK1 and several other proteins to make the repressive complex that depresses BMAL1-CLOCK activity and alters periodicity of the mammalian clock which alters its accuracy. Remember all circadian clocks are flow meters for entropy in a cell.
Solar light input into mammal TTFLs begins with dedicated non visual photoreceptors that elicit signaling that acts to induce (in fungi and mammals) or reduce (insects) the amounts of negative arm proteins mentioned above. In broad outline, Output occurs when the positive Arm heterodimer binds to DNA and activates expression of genes whose products do not impact the TTFL. The key take away is the clock gene actions are PROXIMAL to DNA translation and gene activation. This tells you that light inputs controls gene expression in mammals and it is not the other way around. Altering your genome will not improve your illness or disease if the light and dark environment is repair first.
HOW DOES TIME STAMPING WORK BY LIGHT AND DARK WORK?
Once enough modified protein products accumulate in the cytoplasm of a cell, they are transported into the nucleus where they inhibit the positive element from the promoter to stop transcription of clock genes. The clock gene is thus transcribed at low levels until its protein products are degraded, allowing for positive regulatory elements to bind to the promoter and restart DNA transcription. The negative feedback loop of the TTFL has multiple properties important for the cellular circadian clock. First, it results in daily rhythms in both gene transcription and protein abundance and size, caused by the delay between translation and negative regulation of the gene. The cycle's period, or time required to complete one cycle, remains consistent in each individual and, barring mutation, is typically near 24 hours. This enables stable entrainment to the 24 hour light-dark cycle that Earth experiences from the sun & moon.
Additionally, the protein products of clock genes control downstream genes that are not part of the feedback loop, allowing clock genes to create daily rhythms in other processes, such as metabolism, within the organism. Light and dark cycles are the decentralized controllers of the TTFL.
THE TTFL USES MELANIN TO ELECTROCHEMICALLY TIME STAMP YOUR CELLS. This occurs in the retinohypothalamic pathways anterior to the SCN and it modifies what the SCN signals to all the other molecular clock genes it links to in tissues.

WHY IS MORNING LIGHT SO CRITICAL TO GET RIGHT?
CSP-1 (conidial separation 1) is a morning induced transcriptional repressor with a phosphorylation gated half-life is a key cog in driving EVENING gene expression in mammals. If you do not get AM sun your evening genomic expression will be AWRY. People have forgotten that leptin is released by fat cells and can only enter the hypothalamus under darkness after 4 hours. This should happen at night time. It cannot happen when CSP-1 is not created by AM light. These are the new recent insights into how circadian clocks in your eye and skin achieve phase-specific gene expression. This is how and why leptin resistance exists.

The negative element of the core circadian feedback loop is the frq or frequency gene. The frequency (frq) gene controls the morning-specific rhythmic transcription of a sense RNA encoding FRQ segment. As a result of this action, a long noncoding antisense RNA, qrf, is rhythmically transcribed in an evening-specific manner. It has been reported in the literature that the qrf rhythm relies on transcriptional interference with frq transcription and that complete suppression of qrf transcription impairs the circadian clock. The biological function of qrf transcription and its impact on the circadian clock are not understood in centralized science because centralized science has no light controls at night in labs.

CSP-1 expression is induced by light and glucose, and this finding suggests a rhythmic coordination of qrf transcription with metabolism. Because it is light and glucose we know POMC, ACTH, and melanin are the key to understanding CSP-1 biology. It also means that artificial light during the day or night is especially toxic when you know this is how the mechanism operates.
There are three type of melanins in humans and only one ACTH in humans. All three are used to time stamp the atomic lattice of cells to create an internal map of space time domains to be accurate measuring sticks for the flow of entropy inside of cells. This links melanin biology to Noether's theorem directly. You have blogs on all these ideas now and it is time for you to link them all to comprend what I have been teaching your for 20 years. Light causes modern diseases.
These 3 melanins are ALL extended heterogeneous biopolymers composed of molecular subunits with ambiguous macromolecular topology to modern centralized science. In the literature, an electrochemical fingerprinting technique has been described for melanin, which suggests that natural melanin pigments which contain indole-based tetramers seem to be always arranged into porphyrin-like domains to capture light and measure it in some way useful to the system.

Spectroscopy and density functional theory calculations suggest that sodium ions undergo occupancy-dependent stepwise insertion into the core of porphyrin-like tetramers in natural melanins at discrete potentials that time stamp the internal atomic lattice that allow it to act like a clock to measure the flow of entropy in the cellular system accurately just using light and dark as the feedback loops. It is fully decentralized because light and dark control this process. One is not more important than the other. A loss of melanin implies a loss of accurate time keeping inside the cell or tissue.

Lastly, in humans, the TTFL is a limit cycle, meaning that it is a closed loop that will return to its fixed trajectory even if it is disturbed by its environment, maintaining the oscillatory path on its fixed 24-hour period. It appears this is only true if the melanin structures it uses on surfaces and endogenously remain intact and are chronically replaced and renovated. If the endogenous electrochemical time stamping mechanism is damaged, the TTFL loses its periodicity and chronic modern disease results without any alteration to the DNA or RNA of a cell. These are diseases do not mimic genetic diseases like Tay Sach's. These diseases are far more common than mutational diseases of DNA which are relatively rare. It means our circadian mechanism is open to the environment, and as such is not subject to calories measurement for metabolism because calories only is useful in closed thermodynamic loops.

CITES
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Dr. Jack Kruse
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