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Title page. * RSV Core Assembly: It ’ s All About Nucleation. Carmen Butan, Giovanni Cardone, Naiqian Cheng, Bernard Heymann, Paul Keller, Dennis Winkler. Laboratory of Structural Biology, NIAMS-NIH. Parvez Lokhandwala, John Purdy, Rebecca Craven.

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  1. Title page * RSV Core Assembly:It’s All About Nucleation Carmen Butan, Giovanni Cardone, Naiqian Cheng, Bernard Heymann, Paul Keller, Dennis Winkler Laboratory of Structural Biology, NIAMS-NIH Parvez Lokhandwala, John Purdy, Rebecca Craven Pennsylvania State University College of Medicine

  2. Retroviruses are highly variable in their capsid architecture, both for a given retrovirus and between different retroviruses. • With HIV, the predominant capsid morphology is biconical; with Rous sarcoma virus, it is irregular polyhedra. Cylindrical tubes with and without end-caps are also produced. • .

  3. Cryo-electron tomogram of Rous sarcoma virus Butan et al. (2008) J. Mol. Biol., 376, 1168-81

  4. Angular, curved and miscellaneous cores

  5. Virions with angular cores have many spikes: Virions with tubular cores have few spikes

  6. In retroviruses, the capsid assembles de novo inside the provirion, post-budding Not all of the capsid protein assembles: > 50% remains unassembled (Prevelige et al; Fuller et al)

  7. Measure #Gag ( total # CA) from virion diameter Angular cores correlate with high CA assembly efficiencies: Tubular cores correlate with low CA assembly efficiencies

  8. Observations • The range of polymorphic variation is very large • tubular cores and many “curved”cores - including biconical RSV • cores - fail to package RNP • Hypothesis: a viable core is one that is closed and has successfully packaged the genome(s) and the polymerase

  9. Observations • The range of polymorphic variation is very large • tubular cores and many “curved”cores - including biconical RSV • cores - fail to package RNP • Hypothesis: a viable core is one that is closed and has successfully packaged the genome(s) and the polymerase • Inference: Initiation is key to capsid assembly

  10. Critical concentrations for assembly conc’n of protein subunits in capsids Total conc’n of protein subunits

  11. Nucleation of angular nucleocapsid assembly : a model

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