Lecture 3: Sizing Up Viruses Phage in addition to the Origins of Molecular Biology Views of the Lambda Phage Genome Phage T4 Genome
Brillhart, Aaron, Host; Meteorologist has reference to this Academic Journal, PHwiki organized this Journal The Parts List of a Virus Lecture 3: Sizing Up Viruses http://faculty.washington.edu/jais/microscopy.html Cells in addition to the Viruses That Infect Them http://faculty.washington.edu/jais/microscopy.html +HIV +phage Bacteria T cells Note: the viral parts list is very small (10-100 pieces) with genomes roughly of 10kb. They accomplish so much with so little. Phage in addition to the Origins of Molecular Biology Adapted from Molecular Expressions (Wertz) Analogy: Put 500m of Golden Gate Bridge suspension cable in the back of a FedEx Truck. An infected bacterium: note that virus injects DNA in addition to leaves body behind.
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Views of the Lambda Phage Genome Phage T4 Genome Despite their supposedly simplicity in addition to the existence of sequenced genomes, there is still much that we dont know about what goes on in these genomes. 168,903 base pairs in the genome Life Cycle of a Bacteriophage Rate of packing: 100bp/sec Some assembly required Self-assembly Rate of ejection: 100 – 10000bp/sec Construct a physical model of these processes. Forceful ejection
Who Are the Real Predators Out There The density of bacteriophage in the ocean is 50 x 10^6/ml while the density of sharks is much less than 10^-6/ml! Hendrix et al. It is estimated that 40% of the bacterial background is infected everyday by bacteriophage – that is a predator! Phage as Model Systems as long as Physical Biology Phage provide a setting within which we can ask sharp, quantitative questions in addition to test precise models of biological phenomena. Some universal biological themes: macromolecular assembly in a crowded environment, orchestration in space in addition to time, polymer translocation, gene expression, evolution, natures nanotechnology etc. Close to having a full census (molecular inventory) in addition to full structure of phage. An opinion: phage have not outlived their ability to teach! Rossmann et al. Plant Viruses in addition to the Reconstitution of Life 1955 Fraenkel-Conrat in addition to Williams – first example of complex biological entity reconstituted outside off cell. > 2000 protein units co-assemble with RNA molecule. Headlines: Life created in test tube!
Plant Viruses in addition to the Reconstitution of Life Cowpea chlorotic mottle virus – plant virus. 4 separate RNA molecules constitute its genome. Structure known with atomic resolution. Reconstitution of CCMV Structure of Viruses (Baker et al.) Characteristic size scale is 30-100 nm. Structures are known at atomic resolution – see Viper website. Highly symmetric – think hard about what this implies about assembly!
The HIV Life Cycle Key point: viruses hijack the host cell to make new viruses. Once again, many of the great themes of biology are played out in this microcosm. HIV a convenient in addition to intriguing prism through which to view all of biology. There are many interesting physical processes that take place in this life cycle that will catch our fancy: binding, membrane fusion, transcription in addition to its control, integration, assembly, budding! This one picture literally provides a road map as long as the entire course. Almost all of our big themes are revealed right here! Views of the HIV Genome http://www.gladstone.ucsf.edu/gladstone/html/connections/fig1.html Making a Virus: Mature Virions Fuller et al. Cryo electron microscopy provides a window on virion structure.
Structural Heterogeneity of HIV Particles Each picture is 160nm wide. Benjamin et al. Mean diameter approximately 120nm. Average volume approximately 45,000 nm^3 Approximate mass per virus particle is 1fg (650 Mda) The HIV Protein Parts List This is a nice in addition to thought provoking picture, but Be careful, which features of this cartoon do we REALLY know When shown cartoons like this, you have to ask yourself what is known in addition to how. Compare the cartoon to the actual data – what is the data Try to make a cartoon sometime in addition to it will reveal lots about what we dont know. (Turner in addition to Summers) Making a Virus Fuller et al. Yogi Berra said You can observe a lot just by watching. We will pursue a corrolary: you can learn a lot just by estimating. Estimation question: how many Gag proteins does it take (roughly) to make an HIV virion We begin by considering the immature virus. Does the cartoon make sense To figure out the number of Gag proteins, we need an estimate of the area per protein.
Important Yogi Berra remarks “Nobody goes there anymore; it’s too crowded. “If you come to a as long as k in the road, take it. “You should always go to other people’s funerals; otherwise, they won’t come to yours. “It was impossible to get a conversation going; everybody was talking too much. “You’ve got to be very careful if you don’t know where you’re going, because you might not get there.” Acquiring a Membrane: How HIV Gets Its Lipids This scanning electron microscope picture demonstrates HIV budding (arrows) from the surface of an infected T-lymphocyte magnified 80,000X. http://www.homepage.montana.edu/~spincus/sp-hiv.html http://faculty.washington.edu/jais/microscopy.html Lipid Composition of the HIV Membrane Brugger et al. Fact of life: lipids come in many different shapes in addition to chemical as long as ms. HIV lipid census provides a partial picture of the virion.
What Are Lipids Like Typical sizes: 2.5nm Typical masses: 750 Da Area per molecule: 1/4 nm^2 A useful place to find stuff out is Avanti Polar Lipids. Lipid Content of HIV Particles The actual molecular census of the HIV virus. Capsid Structure Fascinating structure of the internal capsid of HIV. Shape conferred by geometric rules about 5-fold defects. Sundquist et al.
Distribution of Spike Complexes Zhu et al., Nature 2006 Distribution of Spike Proteins Zhu et al., Nature 2006 Viruses in addition to the Great Themes of Biology How do viruses make new viruses How do viruses evolve (including to evade the host defenses) How do cells defend against viruses How do cells make decisions in addition to how is that decision making altered by the presence of viruses Korber et al.
Brillhart, Aaron Host; Meteorologist
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