MOLECULAR BIOLOGY I: NUCLEIC ACID METABOLISM

SC/BIOL 3110, 2020 S1

Bonus Lecture: Last recap

1
Course Announcements:

• Just a reminder: today (Jun 23) is the last day to submit an online course
evaluation. You can use the following URL to complete the evaluation.

http://courseevaluations.yorku.ca/

• Also, final exam is this Friday (Jun 26) from 7 to 10 pm.

• The final exam format will be similar to the midterms. Make sure you have a
calculator ready.

• ~ 65% of the final exam will focus on the last 3rd of the course.

• I will not be using proctortrack because it is not possible to ensure all students
have webcams in such a short time; however, that means I will be using the
same sequential exam question method as before.

• I will post more details on the Moodle page (e.g. how many questions in MC vs.
short answer sections and the marks distribution of the sections) by Friday
morning.

2
Recap of last lecture:

Histones methylation:

• A number of lysines on histone proteins are methylated.

• Note: individual lysines can either be methylated, acetylated or ubiquitylated, but

each lysine can only be modified by one type of PTM at a time.

• Also note that arginines on histones can also be methylated à only mono- or di-
methylated
3
Recap of last lecture:

Position Effect Variegation (PEV):

• See https://smallscienceworks.com/2015/01/28/fifty-shades-of-red-the-superhot-
history-of-pev/ for blog post explaining the history of position effect variegation, and
the identification of Su(var) genes and pathway leading to heterochromatin
formation.

• Started with Muller using X-

rays to mutagenize flies
and cataloguing various
phenotypes, including
red/white/variegated eyes.

• Others performed additional

forward genetic screens to
search for Suppressor and
Enhancer mutations.

4
Recap of last lecture:

Position Effect Variegation (PEV):

5
Recap of last lecture:

Position Effect Variegation (PEV):

• Very simplistically: Su(var) genes code for proteins that help make heterochromatin
whereas E(var) genes code for proteins that antagonize heterochromatin formation

6
Recap of last lecture:

Constitutive heterochromatin formation (simplified):

Step 1: Establishment of Step 2: Recruitment of HP1 Step 3: Propagation of

H3K9-methylation (via it’s chromodomain) heterochromatin

Su(var) 2-1 = HDAC1

Su(var) 3-9 = H3K9 HMT

Su(var) 2-5 = HP1

7
Recap of last lecture:

Constitutive heterochromatin formation:

• In mouse, binding of HP1 brings in more SUV39H1 (via chromoshadow domain) to

methylate neighbouring nucleosomes (i.e. co-operativity) à spreading of
heterochromatin.

• Spreading of heterochromatin is stochastic (i.e. no fixed end site) unless stopped by

defined boundary element (e.g. insulator).

8
Recap of last lecture:

Differential H3methylation marks distinct chromatin domains:

constitutive facultative
euchromatin heterochromatin heterochromatin

K4-methylated H3 K9-methylated H3 K27-methylated H3

9
 Recap of last lecture:

Differential histone methylation form distinct types of chromatin:

• Different methylated residues on histone H3 and H4 and methyl-H3/H4 binding

proteins:

Trx/MLL = H3K4 HMT

Su(var) 3-9 = H3K9 HMT
E(Z)/EZH2 = H3K27 HMT

SET domain-
containing proteins
10
Recap of last lecture:

Histone code paradigm:

• Variety of inhibitors of histone modification-writers/erasers/readers have been developed

and used in clinical trials for different diseases such as different types of cancer.

11
Recap of last lecture:

Using/developing epigenetic drugs:

• Many small molecule inhibitors designed to target DNA methylation or histone code
writers/erasers/readers are currently being developed and tested against various
diseases such as cancer.

e.g. HDAC inhibitors (SAHA)

histone methyltransferase inhibitors
Brd4 inhibitors

12
 Recap of last lecture:

Brd4 – Ac-H4 reader important for transcriptional activation:

• Brd4 contains tandem bromodomains that bind to acetylated-H4. Binding of Brd4 to

regulatory elements (e.g. enhancers/promoters) in turn recruit TFs and other factors
(e.g. PTEFb) that promote/facilitate transcription elongation.

BET family members: all have double BDs

and all inhibitied by JQ1

13
Epigenetics and chromatin regulation:

Potential therapeutic uses of the Brd4 inhibitor JQ1 (and similar compounds)?

BRDT
Other uses?

14