Font Sizes: A A A

Molecular Cell Biology I

Faculty of Medicine
6 credit points, terminal examination, Gábor Bánhegyi; AOKOVM464_1A

2016/2017 Spring semester

Teaching secretary: Gergely Keszler

Intro


Description of the curriculum
The principal aim of the course is to provide an insight into the storage and expression of genetic information throughout replication, transcription and translation as well as our current understanding of the regulation of gene expression, followed by an up-to-date summary of current methods in molecular biotechnology.

Module I. DNA, RNA and protein synthesis (storage and expression of genetic information)
Nucleic acids – structure and function. Bases, nucleosides, nucleotides, DNA structure, DNA denaturation, hybridization. DNA replication. Replication in prokaryotes, leading and lagging strand. Okazaki fragments. DNA-dependent DNA polymerases. DNA ligase. Telomerase. topoizomerases. Replication in eukaryotes. Structure of eukaryotic chromosomes. Mitochondrial DNA. Nucleosome structure. DNA repair. Types of DNA damage; mutations, frameshift, nonsense mutations, mismatch repair. Coordination of repair and replication. Transcription in procaryotes. Structure of RNA; t-RNA, r-RNA, m-RNA, differences between the procaryotic and eucaryotic genomes.  Transcription complexes, initiation, elongation, termination in prokaryotes. Transcription in eukaryotes, RNA polymerases, promoters, enhancers, silencers. Transcription factors. Processing of mRNA, mechanism of splicing. Alternative splicing, mRNA editing. The genetic code. Activation of tRNA. Mechanism of translation, initiation, elongation, termination. Antibiotics. Posttranslational modification of proteins. Protein folding, sorting, quality control and transport into intracellular compartments. Ubiquitination and intracellular proteolysis. Epigenetic regulation of gene expression. Mobile genetic elements and the molecular biology of viruses.

Regulation of gene expression in prokaryotes. The operon model. Positive and  negative regulation in the lac operon. Regulation of gene expression in eukaryotes at the transcriptional  level. Role of chromatine structure; covalent and non-covalent chromatin modifying activities and DNA methylation (epigenetics).  Post-transcriptional regulation in eukaryotes. Regulation of  mRNA stability; microRNAs. Translational regulation.

Module II. Methods in molecular biology and gene technology
Principles of recombinant DNA technology: molecular cloning, restriction endonucleases. Genomic and cDNA libraries. Blotting techniques (Southern, Northern, Western) and their utilization. DNA microarrays. PCR and its application  in molecular biology. Recombinant vectors (reporter and expression vectors); synthesis of recombinant proteins. Transgenic, knock-out and knock-in animals in medical research. Human gene therapy. The Human Genome Project and its results: organization and polymorphic nature of the human genome; implications for human traits and diseases. Genotyping methods (PCR-RFLP, PCR-ASA). Application of bioinformatics and systems biology in biological and medical research.

Requirements for acknowledgement of the semester

  1. Participation in the laboratory practicals, consultations and seminars is obligatory; students have to sign the attendance sheets at the end of every lesson. In case of more than three absences from the practical lessons for any reason, the semester will not be acknowledged and the student is not allowed to sit for the semifinal exam. Missed practicals can be completed only in the same week at another group; certificate from the host teacher should be presented by the student to his/her own teacher. The schedule of practical lessons, consultations and seminars can be downloaded from the official homepage of the Institute.
  2. The oral midterm examination (week 8) and the written test (week 13) have to be passed before the commencement of the examination period. The material of the midterm examination corresponds to that of lectures of weeks 1-7 while the lecture material of weeks 8-12 will be asked ont he written test. Failed midterms might be retaken twice.

Semifinal exam
The course is finished with an oral exam. Students will be examined by a two-member examination committee. 4 topics taken from the topic list have to be answered.

Recommended textbooks
Lodish: Molecular Cell Biology (8th edition)
Hrabák: Laboratory Manual – Medical Chemistry and Biochemistry

Lectures


Location: EOK (Tűzoltó u. 37-47.) Szent-Györgyi Lecture Hall
Duration: 55/60 min (Wednesdays 16:10-17:05 & Thursdays 14:50-15:50)
WeekDateTopicLecturer
1.30 Jan - 3 FebEucaryotic and procaryotic cell, the genetic informationKeszler
Basic concepts of molecular cell biology, structure and function of nucleic acidsKeszler
2.6-10 FebThe chromosomes and the organization of DNA IKeszler
The chromosomes and the organization of DNA IIKeszler
3.13-17 FebThe replication, repair and recombination of DNA IRónai
The replication, repair and recombination of DNA IIRónai
4.20-24 FebTranscription ICsala
Transcription IICsala
5.27 Feb - 3 MarTranscription IIICsala
The regulation of gene expression, nuclear receptorsSipeki
6.6-10 MarTranscriptional factors, DNA-binding domainsSipeki
The genetic code and translationMészáros T.
7.13-17 MarPosttranslational modification of proteinsMészáros T.
8.20-24 MarFolding and quality controlMészáros T.
Proteostasis, the ubiquitin-proteasome systemSőti
9.27-31 MarThe types of autophagySőti
Mobile genetic elements, virusesCsala
10.3-7 AprMobile genetic elements, virusesCsala
Evolution of genes and the genomeNemoda
11.18-21 AprEpigeneticsNemoda
Methods in molecular biology IRónai
12.24-28 AprMethods in molecular biology IIRónai
13.2-5 MayMethods in molecular biology IIIRónai
Methods in molecular biology IVRónai
14.8-12 MayBioinformatics, systems biology IKapuy/Csermely
Bioinformatics, systems biology IIKapuy/Csermely

Labs/Seminars


Location: EOK (Tűzoltó u. 37-47.) 1st floor, corridor ‘D’
WeekDateLab / Consultation
1.30 Jan - 3 FebProteins I
2.6-10 FebProteins II
3.13-17 FebPaper and thin layer chromatography*
4.20-24 FebGel electrophoresis*
5.27 Feb - 3 MarDetermination of urease activity
6.6-10 Mar
(Friday between 11.00-13.00 is off)
β-Galactosidase
7.13-17 Mar
(Wednesday: holiday)
Molecular biology (Consultation)
8.20-24 MarMidterm (oral)
9.27-31 MarPlasmid digestion*
10.3-7 AprPCR-based genotyping I*
11.18-21 Apr
(Monday is off)
PCR-based genotyping II*
12.24-28 Apr
(Wednesday is off)
Biotechnology (Consultation)
13.2-5 May
(Mondayis off)
Written test
14.8-12 MayGel filtration

*The order of marked labs varies from group to group. See the detailed schedule.

Groups


GroupTeacherDayTime
EM/1SipekiTuesday10:45-14:00
EM/2VereczkeiTuesday10:45-14:00
EM/3Mészáros T.Tuesday10:45-14:00
EM/4SasváriTuesday10:45-14:00
EM/5SzeitnerMonday15:15-18:30
EM/6KeszlerMonday11:45-15:00
EM/7BőgelMonday11:45-15:00
EM/8TányaMonday15:15-18:30
EM/9KardonMonday11:45-15:00
EM/10LizákMonday11:45-15:00
EM/11MargittaiMonday15:15-18:30
EM/12HrabákFriday11:30-14:45
EM/13CsalaMonday13:20-16:35
EM/14StroeMonday15:15-18:30

Topics


Coming soon.

Midterm I


Topics for the first midterm examination

  1. Comparison of pro- and eukaryotic cells: compartmentation and the role of the most important subcellular organelles
  2. The chemical structure of nucleotides. The primary structure of nucleic acids (DNA and RNAs)
  3. Condensation of the genetic material in pro- and eukaryotic cells. The role of topoisomerases and chromatin proteins
  4. The structure of human chromosomes and its alterations during the cell cycle
  5. The structure of the human genome: coding and gene regulatory sequences. Non-coding genomic sequences: introns, pseudogenes, repetitive sequences
  6. Principles of the semiconservative DNA replication: replication fork, leading and lagging strand synthesis
  7. Comparison of DNA replication in pro- and eukaryotic cells: principles, enzymes, proteins
  8. Telomeric repeat sequences and their maintenance
  9. Classification of DNA damages and repair systems. Repair of base deamination
  10. Formation and repair of thymine dimers. Mismatch repair
  11. Classification of point mutations. The origin of spontaneous point mutations. DNA polymorphisms. Possible effects of DNA sequence alterations on the corresponding mRNA and proteins 
  12. Structure and function of the prokaryotic RNA polymerase holoenzyme. Transcriptional initiation and termination in bacteria; the prokaryotic transcription unit
  13. Types of RNA and their biological importance. Synthesis and maturation of rRNAs and tRNAs
  14. Regulation of transcription in prokaryotes. Strong and weak promoters, constitutive genes, operons, positive and negative regulation
  15. Structure of eukaryotic genes. Transcriptional initiation and termination in eukaryotic cells
  16. Regulation of transcription in eukaryotes. Specific transcription factors, cis- and trans-acting regulatory sequences, coactivators, corepressors
  17. Processing of eukaryotic mRNAs
  18. Regulation of eukaryotic gene expression at post-transcriptional levels
  19. Post-transcriptional modification of eukaryotic mRNAs and regulation of their stability
  20. DNA-binding proteins and their characteristic structural elements in prokaryotes (helix-turn-helix) and eukaryotes (histone fold, helix-turn-helix, zinc finger, leucine zipper) with examples
  21. Structure and function of nuclear receptors (steroid, thyroid, aryl hydrocarbon receptor)
  22. The genetic code. Structure and function of tRNAs, aminoacyl-tRNA synthetases, the codon-anticodon hybridisation
  23. The structure of prokaryotic and eukaryotic ribosomes, the ribosome cycle, tRNA binding to ribosomes
  24. Initiation of translation in pro- and eukaryotic cells. Regulation of eukaryotic translation via phosphorylation of eIF2
  25. Elongation and termination of pro- and eukaryotic translation. Pharmacological inhibitors of translation

Download


Lecture slides, seminar handouts, laboratory manuals are available here.
The password to download pages can be obtained from the laboratory teachers personally.