Clock genes Flashcards

1
Q

How many clock genes are present between species?

A

Many many many

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2
Q

True/False? Even though clock genes have different names, their functions are all similar

A

True

“the actors are different but the plot is the same”

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3
Q

What are the two conserved features of clock genes?

A

Oscillating mRNAs, proteins or activity

Autoregulatory feedback loops

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4
Q

Describe the basic mammalian clock and when Period protein is expressed the most

A

CLOCK and BMAL1 proteins associate and activate transcription of Per1,2,3 and Cry1,2, PER1,2,3 and CRY1,2 proteins inhibit CLOCK-BMAL1 complex leading to inhibition of Per123/cry12 transcription

Per1 RNA is most concentrated during the day, and PER protein is most expressed at CT 12`

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5
Q

Why is CLOCK/BMAL1 called a negative autoregulatory loop?

A

Because CRY/PER INHIBIT further synthesis

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6
Q

How do drosophila differ from mammals in terms of their clock genes?

A

CLOCK-CYCLE instead of CLOCK-BMAL1

just PER instead of PER1,2,3
TIM instead of CRY1,2

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7
Q

How was the period gene determined to affect circadian rhythm in drosophila?

A

“period mutants” would hatch at irregular periods

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8
Q

Can Per function without Tim in drosophila?

A

No, Per needs Tim to enter the nucleus and to be stabilized

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9
Q

What is interesting about the PAS domain in the mammalian clock gene?

A

It shares homology with Per (in drosophila)

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10
Q

How was clock deduced to be a transcription factor?

A

Q-rich region, a hallmark of many TFs

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11
Q

Do humans have Per gene?

A

Yes! 3 of them

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12
Q

How are Per genes controlled?

A

Via CLOCK-BMAL1 complex

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13
Q

What are cryptochromes?

A

A group of light-sensitive flavoproteins (FAD binding proteins), similar to photolyases (light-sensitive DNA repair)

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14
Q

Are Cryptochromes transcription factors?

A

Yes and no

They don’t bind DNA themselves, but they bind to Transcription Factors and prevent them from activating transcription

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15
Q

What happens if Cry1 is knocked out? Cry2? Both?

A

Cry1 KO: FRP is shorter
Cry2 KO: FRP is longer

Both: No clock present

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16
Q

True/False? Activity of a protein is roughly proportional to the expression of its gene

A

FALSE

17
Q

How is REV-ERBalpha involved in circadian rhythms?

A

Rhythmically represses Bmal1 gene and its absence alters circadian rhythms

18
Q

How is REV-ERBalpha expression regulated?

A

By BMAL1-CLOCK

19
Q

What is ROR protein and?

A

Activates Bmal1 gene and opposes REV-ERB

20
Q

Is there more to it than what we’ve seen in class?

A

Yes, lost more proteins and enzymes and co-activators etc

21
Q

Of the crazy list of other proteins seen in class, what else should we know about?

A

BMAL2 (like BMAL1) and NPAS2 (like CLOCK)

22
Q

What’s another layer of complexity on top of the typical mammalian rhythm seen in class?

A

Post-translational modifications

23
Q

What is the most common Post translational modification? Second most?

What do they do?

A

Phosphorylation (regulation of clock protein stability, activity, intra-cellular localization)
Ubiquitination (Regulation of clock protein stability and more)

24
Q

What enzyme phosphorylates most circadian rhythm proteins?

A

Casein Kinase 1 epsilon

Affects how much Per is degraded and how fast they can transit in/out of nucleus

25
Q

What is the purpose of Glycogene Synthase Kinase 3 beta (GSK3B)?

A

Phosphorylation and stabilization of REV-ERB alpha

26
Q

How does lithium affect circadian rhythms?

A

Li inhibits GSK3B, thus preventing REV-ERB alpha from being phosphorylated, and is thus degraded

27
Q

How does Casein Kinase 1 Delta/Epsilon lead to degredation of PER 1/2 in mammals?

A

CK1 phosphorylates PER, Phosphorylated PER is then ubiquilated by B-TRCP1/2, leading to proteasomal degradation