[PDF] Untitled 02-Feb-2013 Flavin Adenine

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PRE CLASS SURVEY

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Have you ever

participated in research? (25 seconds)

A. YES

B. NO

Why do you get sleepy?

(25 seconds)

A. Run out of ATP

B. Epinephrine Hormone

C. Melatonin Hormone

D. Sleep Fairy

Do you know what the

circadian rhythm is? (25 seconds)

A. YES

B. NO

What time do you

normally go to sleep? (25 seconds)

A. Before 10

B. Between 10 and 12

C. Between 12 and 2

D. Between 2 and 3

E. After 3

End of the quiz

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Outline

`Circadian Rhythms `Circadian Loop `Cryptochrome `Research

What are circadian rhythms?

`Latin roots `Circa --around `Diem ²aday `A roughly 24 hour cycle controlling physiological processes `Found in most living things `Plants, Animals, Fungi and Cyanobacteria `Self Sustained cycle but directed by exogenous (external) cues `Zeitgebers²´PLPH JLYHUVµ

Zeitgebers

Jürgen Aschoff(1913-98)

Light

Temperature

Social Interactions

Pharmacological

Manipulation

Physical Activity

Eating and

Drinking Patterns

Suprachiasmatic nucleus

`Small, grain of rice sized, region of hypothalamus in the brain `Consists of only 20,000 neurons `Uses light/dark cycles for entrainment `Melanopsinphotopigment `´PMVPHU ŃORŃNµ IRU synchronizing all of the circadian clocks within the body

ILJOP·V HIIHŃP RQ FLUŃMGLMQ 5O\POPV

Abnormal Circadian Rhythms

What happens if the circadian rhythms are disrupted??? `Jet Lag `Shift Work Sleep Disorder `Obesity `Type 2 Diabetes `Increased Risk of Cancer `Seasonal Affective Disorder `Depression

Circadian Sleep Disorders

`Delayed Sleep Phase

Disorder

`Familial Advanced

Sleep Phase Syndrome

`Non-24-hour Sleep-

Wake Syndrome

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Question 1. (70 seconds)

`7OH ŃLUŃMGLMQ ´PMVPHU ŃORŃNµ POMP V\QŃOURQL]HV POH peripheral clocks in the body is located within the brain of humans. Which part of the brain is this and how does it entrain the body to the light day cycle of earth?

`A. SuprachiasmaticNucleus; Cryptochrome `B. SuprachiasmaticNucleus; Melanopsin `C. Pituitary Gland; Melatonin Hormone `D. Pineal Gland; Cryptochrome Protein `E. Pineal Gland; Melanopsin

Answer

`7OH ŃLUŃMGLMQ ´PMVPHU ŃORŃNµ POMP V\QŃOURQL]HV the peripheral clocks in the body is located within the brain of humans. Which part of the brain is this and how does it entrain the body to the light day cycle of earth? `A. SuprachiasmaticNucleus; Cryptochrome `B. SuprachiasmaticNucleus; Melanopsin `C. Pituitary Gland; Melatonin Hormone `D. Pineal Gland; Cryptochrome Protein `E. Pineal Gland; Melanopsin

Question 2 (60 seconds)

`You have just obtained a sample of organisms from the thermal vents at the bottom of the ocean. You discover that they do not have circadian rhythm. What can you say from this observation? `A. Circadian clock components always existed `B. Circadian clock components evolved in the deep ocean `C. Circadian clock components evolved surface of the ocean or on land `GB FLUŃMGLMQ ŃORŃN ŃRPSRQHQPV MUHQ·P UHMO `E. None of the above

Answer

`You have just obtained a sample of organisms from the thermal vents at the bottom of the ocean. You discover that they do not have circadian rhythm. What can you say from this observation? `A. Circadian clock components always existed `B. Circadian clock components evolved in the deep ocean `C. Circadian clock components evolved surface of the ocean or on land `GB FLUŃMGLMQ ŃORŃN ŃRPSRQHQPV MUHQ·P UHMO `E. None of the above

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2NM\ VR RH·YH OHMUQHG M

little bit about circadian rhythms but how do they work at molecular level??

History of Circadian Loop

`period gene first discovered in

Drosophila

`1984 ²period gene cloned independently by Young and Rosbash labs `Large protein of more than 1,200 amino acids `PER-like immunoreactivitywas shown in different orders of animal species suggesting a functionally conserved clock throughout the animal kingdom `Period (PER) `Timeless (TIM) `Circadian LocomotorOutput

Cycles Kaput (CLOCK)

`Cycle (CYC)

Circadian Genes

Mammalian Orthologs

`Period AEPeriod `Timeless AECryptochrome (CRY) `CLOCK AECLOCK `CYCLE AEBMAL-1/ARNTL `ARNTL: Aryl hydrocarbon receptor nuclear translocator-like, `BMAL: brain and muscle aryl hydrocarbon receptor nuclear translocator(ARNT)-like

CLOCK and BMAL1

`Transcriptional factors which contain Basic Helix-Loop-Helix (bHLH) DNA binding domain `Bind to consensus sequence called E-Box `E-Box is at the promoter region of many circadian genes including PER and CRY

PER AND CRY

`mRNA exits nucleus and is translated to form PER and

CRY proteins in the cytosol

`PER and CRY dimerizeand enter nucleus `Inhibit CLOCK-BMAL1 transcription

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QUIZ `GET READY!!! `GET YOUR I>CLICKERS OUT AND TURN THEM ON!!!

Question 3 (45 seconds)

`Which is a negative inhibitor in the circadian loop? `A. ARNTL `B. CLOCK `C. PER/CRY `D. E-Box `E. None of the above

Answer

`Which is the negative inhibitor in the circadian loop? `A. ARNTL `B. CLOCK `C. PER/CRY `D. E-Box `E. None of the above

Question 4 (60 seconds)

`Which structural components make up the area of the CLOCK/BMAL1 heterodimerthat binds to the E-box region of the period and cryptochrome gene promoters? `A. RossmanFolds `B. Beta barrels `C. Anti-Parallel ɴ-sheets `D. Beta sandwich `E. ɲ-helixand loops

Answer

`Which structural components make up the area of the

CLOCK/BMAL1 heterodimerthat binds to the E-box

region of the period and crytochromegene promoters? `E. ɲ-helixes and loops

End of the quiz

Turn OFF your

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Cryptochromes

`Flavoprotein superfamily `Photolyase homology region (PHR) `FAD cofactor `C-terminal discrepancies

Drosophilacryptochrome

FlavinAdenine Dinucleotide

`Also participates in oxydative phosphorylation and citric acid cycle `Involved in electron transfer

Magnetoception

`Cryptochromesin the photoreceptor neurons of birds' eyes are involved in magnetic orientation during migration `Cryptochromesare also essential for the light- dependent ability of

Drosophila to sense

magnetic fields.

FU\SPRŃOURPH·VMetabolic Role

`CRY, modulates gluconeogenesis through interference with glucagon and inhibition of cyclic AMP signaling `Glucocorticoidregulation of the circadian clock modulates glucose homeostasis

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Chemistry 3332 Instructor: Scott Rush 21/2/13

If we have FAD in the cryptochrome what

kind of function will you expect? `A. Methylation `B. Electron Transfer `C. Glycosylation `D. Substrate level phosphorylation `E. None of the above

Question5 (60 seconds)

If we have FAD in the cryptochrome what

kind of function will you expect? `A. Methylation `B. Electron Transfer `C. Glycosylation `D. Substrate level phosphorylation `E. None of the above

Answer

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