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Viruses. Objective: Explain reverse transcription and how retroviruses use it during infection, as well as how it is used in biotechnology. Task: Please copy two key definitions into your notes: Reverse Transcription : making a dsDNA molecule from an RNA template
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Viruses Objective: Explain reverse transcription and how retroviruses use it during infection, as well as how it is used in biotechnology. Task: Please copy two key definitions into your notes: Reverse Transcription: making a dsDNA molecule from an RNA template Gene splicing: inserting DNA from one organism into another. The result is a transgenic organism
Ebola Zaire • Most lethal viral infection known to science ( > 85% of cases fatal) • Antisense ssRNA virus (ss = singlestranded) • 19 kbp genome (human = 3.1 Gbp) codes for 8 proteins
Today’s Big Ideas • Viruses are small genetic entities that are obligate pathogens. (the only way they can survive is through other cells) They are not cells, and contain no ribosomes. • Most viruses defy the normal rules of DNARNAProtein, • some of them store information as RNA • Some can perform reverse transcription • High diversity and mutation rate • Virus may follow a lytic lifecycle or a lysogenic one • Lytic cycle = makes new viruses directly • Lysogenic cycle = inserts genetic info into host.
Lysogenic Cycle: viral RNA reverse transcribed into host DNAex: HIV, HPV, others
Important Characteristics • RNA virus • H & N refer to different versions of viral proteins. • ~100 nm (0.1 μm) diameter. spherical
Step 1: Hemagglutinin Protein Binds to Epithelial Cell in Nose or Lungs H Protein Lung Cell
Hemagglutinin • http://biology.kenyon.edu/BMB/Chime2/2005/Cerchiara-Holsberry/FRAMES/start.htm
Step 4: vRNA Enters Nucleus, Begins making virus materials • vRNA and vRNA polymerase begin replicating and transcribing the virus genes. • Viral genes are transcribed by host cell ribosomes, producing proteins that take over the cell: • Break down cell mRNAs for parts • Use the nucleotides to copy genetic material • Build proteins with cell ribosomes for new viruses
6 Steps of Viral Infection • Attachment • Endocytosis • Unpackaging • Viral Synthesis • Assembly • Release
Human Immunodeficiency Virus: HIV Do Now: Begin reading the background information about HIV/AIDS. We will complete flu presentations momentarily.
Who HIV Strikes: High Risk Behaviors *Includes persons with a diagnosis of AIDS from the beginning of the epidemic through 2007.**Heterosexual contact with a person known to have, or to be at high risk for, HIV infection.*** Includes hemophilia, blood transfusion, perinatal exposure, and risk not reported or not identified.
RNA Virus • HIV stores its genes in the form of RNA. Each free virus has 2 identical copies of the same molecule.
HIV is a Retrovirus • A retrovirus is a type of virus which inserts its genetic material into the DNA of the host cell it infects. • Because the viral genes become part of cell’s own DNA, it is nearly impossible to completely get rid of an HIV infection. • This is also the reason HIV infection may take 10-15 years to cause AIDS
STEP 1: Gp120 protein on virus attaches to CD4 protein on white blood cell
STEP 2: Reverse Transcriptase makes DNA copy of vRNA vRNA DNA-RNA Hybrid Molecule Reverse Transcriptase
Reverse Transcription: A Powerful Tool for Humanity & Viruses • Remember that transcription is the process by which a cell makes an RNA copy of a gene stored as DNA. The RNA molecule produced provides the instructions for protein synthesis to the ribosomes.
Transcription: Forward and Backwards • Transcription: DNA RNA • Reverse Transcription: RNA DNA • REVERSE TRANSCRIPTION is the process by which a DNA copy of an RNA molecule is made. • The ability to make double stranded DNA from single stranded RNA is how retroviruses insert themselves into host DNA • It is also a key tool of biotechnology, allowing for the study and storage of all genes being expressed in a cell.
STEP 3: Integrase inserts DNA copy of viral genes into host DNA. Integrase protein Virus DNA Host DNA
STEP 4: Proviral DNA is transcribed into mRNA. Host DNA Proviral mRNA proviral DNA
STEP 5: Viral Proteins are made and processed Protease Viral pre-protein Finished viral protein
STEP 6: Viral RNA and proteins assemble into a capsid Protease Viral RNA Reverse Transcriptase
Step 7: New virus buds from cell