BIOL 263: Molecular Biology


Syllabus

Fall Semester 2023

ALWAYS UNDER CONSTRUCTION

October 19, 2023
(Latest update overrides all previous versions, electronic and print.)

MWF 10:10 AM-11:00 AM (Period 3)
TOM 101

Dr. Wade Powell
Proper Nouns: "Wade," "Professor Powell," "Dr. Powell" are all appropriate
Pronouns: he/him/his
FSH 202/204, (740) 427-5396
powellw[a]kenyon.edu

Office hours:




Introduction:

The molecular and genomic basis of life is at the heart of modern biology. In BIOL 263, we will learn techniques and explore research questions at the forefront of molecular biology, focusing on the mechanisms by which the information of the genome is expressed to form the functional molecules of living cells and organisms. The processes of DNA replication, recombination and repair, transcription of RNA from DNA templates, and translation of RNA into protein are discussed in the context of current research, frequently using primary literature. The function of genes and regulation and measurement of gene expression are treated in depth. Students analyze and publish interactive tutorials on the structure and function of key macromolecules. This intermediate-level course presumes a strong background in the basics of protein structure/function, central dogma processes, fundamental molecular techniques for manipulating nucleic acids and proteins, and general chemistry.

Required prerequisites: BIOL 115, 116; one year of chemistry (CHEM 121+124 or CHEM 122); or permission of the instructor.  Organic Chemistry is not required (although many students take this course at the same time).

Learning Goals:


Reading Materials and Resources:



Policies and Expectations:

Assignments and Grades: Your letter grade is assigned at the end of the course based on performance in the following areas:


Important Links:
(Note: Some resources can only be accessed on Kenyon campus.)

Course Schedule


Day
Date
Topic
Primary Reading Assignment
Supporting Reading
Part I: Molecular Interactions
F

Aug 25  

 
Course Introduction. Nucleic acid and nucleotide structure.
 Cox pp. 62-64 and174-184. DNA intro; B-DNA; A-DNA
M
Aug 28
Amino acid structure.  Weak bonds in macromolecules.  Cox pp. 68-77 and 93-98 Iesp. Fig. 4-3)   Weak bonds. Amino Acids; CAP; Ident
W
Aug 30
Advanced DNA structure:  B/A/Z-DNA structures; Base Tautomers, Supercoiling Cox 185-189; 179-180; 304-317; FIg. 19.15 (p. 676) Adv DNA; Z-DNA ; Triplex; Quad; base tautomers; supercoiling; Topoisomerase
F
Sept 1
Protein Structure: Helices, strands, sheets, interactions between aa residues

Cox 102-119

AA2 in proteins.
M
Sept 4

Methods for Solving Macromolecular Structures. Intro to pdb and JMOL. RSCB Protein Data Bank

Protein Purification: Cox 100-101 (Highlight 4-1)

Structure Determination: Cox 120-125 [no CryoEM :( ]

A summary of structural biology approaches

NIH Structure Primer

Protein purification by liquid chromatography

Critical Thinking:
How do you tell if a reported structure is correct?

Biochemistry vs. structural biology

W
Sept 6

Structure of a DNA binding protein, ADAR1


 

Schwartz 99;
ADAR1 structures in rcsbPDB
ADAR activity and consequence


The famous Gillen guide to reading journal articles.
Vocab;


Part II: Gene Expression
F
Sept 8
Transcription initiation, elongation, termination
Cox 519-536  Structures: RNAP open complex; RnaPol-Euk; movies!
M
Sept 11
Transcription Research:  RNA polymerase "scrunching"

Kapanidis et al. 2006

Supporting Info

Cox p. 533 (Fig. 15-15)

Background: Single-molecule techniques explained.

DNA footprinting: Cox 700-701 (Highlight 20.1, Fig. 2)

W
Sept 13

Bacterial Transcription Regulation at level of transcription initiation.

Activators and repressors:Cox 665-675
Operons 693-703
Regulons: 705-707
CAP-CTD; Benoff 1999;
Liu 2017
Lac repressor
Cooperative binding
F
Sept 15
Bacterial transcription regulation, cont'd: Regulation of events after initiation: leader sequences, riboswitches, attenuation Cox 704-705; 707-711
 -Soukup 2004 review
-Breaker Lab Riboswitch site (Yale) &movie
-riboswitch methods/data
M
Sept 18
Unique mechanism of attenuation in Bacillus.
Paper:  Antson 99 trp RNA-binding attenuation protein.

 
W
Sept 20
Catch-up or review

 


F
Sept 22
Exam I (in class) .  
M
Sept 25
Eukaryotic General Transcription Factors. DAB Pol II; TBP; TAF; solving the structure of RNA pol II.
Cox 537-548
Techniques:
gel shifts: 700, FIg. 1 (Highlight 20.1)
chromatography: 100-101(again).
W
Sept. 27
Cox 537-548 PIC assembly structures
PIC assembly movie (download movie from supplementary info link in article)
F
Sept 29
Eukaryotic Basal Transcription Research:


M
Oct 2
Eukaryotic Transcriptional Regulation:  Regulatory elements and transcription factors
Cox 730-737; Highlight 21-2 (738-739); 740-745  
W
Oct 4
Nucleosomes & Transcriptional regulation;  Nucleosome tutorial Cox 331-343; 728-730
Histone binding sites: Nature N&V 2006

 
F
Oct 6
Fall Break; No Class  
M
Oct 9


Research in Eukaryotic Transcriptional Regulation: Chromatin Immunoprecipitation; Histone acetyltransferase recruitment

 

Sharma 2002

See moodle for guided meditation assignment.

ChIP:Watson Ch. 21, pp. 751-752;

cross-link chemistry

PCR: Cox p. 221-226
Remember PCR?

W
Oct 11
Transcript Splicing; Splicing2; Spliceosome Cox 553-565
F
Oct 13
[Wade Away]

M
Oct 16
Splicing conclusion
Translation: Peptide bond formation; ribosome composition
Alternative splicing: Cox 561-562
Peptide bond formation: Cox 65, 644
Ribosome basics: 617-626
Due date: Molecular Tutorial Proposal (instructions and electronic submission page on moodle)
W
Oct 18
Translation basics; initiation. Initiation. PDB; ribosome structural primer Cox 617-626; 626-630; 630-63  
F
Oct 20
Translation elongation, termination. tRNA charging Cox 639-649
M
Oct 23
Translation Research: Ribosome hybrid state Dorner 2006
W Oct 25  RNA Interference
Animation from Nature Reviews Genetics.
 Cox: 681; 774-781.  Wilson and Doudna 2013 (RNAi review paper)
F
Oct 27
Exam II (points to ponder)    
Part III: DNA Replication, Repair, Recombination
M Oct 30 DNA Replication; Beta ring ; DNA Rep; Replisome
Web Project Workshop, 7-9 PM;
Cox 363-376
W
Nov 1
DNA Replication, cont'd.;
Web Project Workshop, 7-9 PM;
Cox 377-393; 398
 
R Nov 2
Web Project Workshop, 7-9 PM; 

F Nov 3 Replication Paper--Single-molecule replisome study:
Are leading and lagging strand polymerases really coordinated?
Graham et al. 2017
Guided meditation on moodle due at beginning of class
Don't ignore the supplemental information figures!
Sun Nov 5

Web Project Workshop, 7-9 PM;

   
M
Nov 6
DNA Mutation and Repair
Cox 413-445

W
Nov 8
DNA Repair Research


F
Nov 10
Homologous Recombination;
Cox 449-466 Experiments; RuvA; PNAS; Tn;
M
Nov 13
Telomerase and telomeres. Telomerase structure. Cox 399-405
Blackburn 2000 review

W
Nov 15
Paper:  Ter, tert, and cancer Li et al 2004
F
Nov 18

Generation of knockout mice by homologous recombination


 
M-F Nov 20-24
Thanksgiving Break; no class


M
Nov 27
CRISPR Genome Editing Cox 246-248 (esp. Fig. 7-33)

W
Nov 29
Paper:  Application of CRISPR to characterize lncRNA function Espinosa 2016 (News & Views piece)
Paralkar 2016 (research article)

F
Dec 1
Exam III    
M
Dec 4 
In-class pdb work time    
W
Dec 6
Student Project Presentations . Completed JMol Tutorial
F
Dec 8
Student Project Presentations

.

 
R
Dec 14
Revised Molecular Tutorials Due by 4:30 PM (corresponds with end of scheduled exam period) Final Submission (via moodle)

 
     

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