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Bio 430 SEPA-1 Presentation

Bio 430 SEPA-1 Presentation - hang Y et al 2009 Cell Volume...

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hang, Y. et al. 2009. Cell Volume oanna Agwa and Mariam Mahroos SEPA 1 in the Mediation of  Recognition and Degradation of P  Granule components by Autophagy
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Outline
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P granules
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P granules What are P granules? Accumulate in germ line cells but not somatic cells (Strome and Wood, 1981) Oocyte derived cytoplasmic components, possible role in germ line specification (Pitt, 2001)
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Autophagy: The Machinery Atg1 P Atg 8 and Atg 12 Atg6/Vps34 Atg9 Klionsky, 2005 and Suzuki, K; and Ohsumi, Y. 2007
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Autophagy: The Physiological significance
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Physiological importance: Removal of  aggregated proteins What is known? p62 may link protein aggregates to autophagy Possibly via interaction with Atg8 (Bjorkoy et al., 2005; Pankiv et al., 2007) What is unknown? The process for depletion of P granule aggregates in somatic cells
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The Purpose of the Study
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1.) How they form in somatic cells
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1A.) Formation of P granules in absence of  autophagy Results: Inactivation of lgg-1 leads to accumulation of PGL-1 granules in somatic cells Staining with K76 determined dispersal of granules in somatic cell cytoplasm
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1A.) Formation of P granules in absence of  autophagy Results: Co-staining with anti-PGL-1 and anti-PGL-3 showed both P granule components co-localized to granules in somatic cells Other autophagy mutants also showed formation of granules in somatic cells
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1A.) Formation of P granules in absence of  autophagy Conclusion: Autophagy prevents formation of P granules in somatic cells.
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Traces of P Granule components found in  membrane bound somatic cells Methods Immunoelectron microscopy Incubated sections of early stage wild-type embryos with k76 Then incubated with gold-conjugated secondary anti-body Results Gold particles only rarely observed EM sections found PGL in 9 membrane bound structures
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Traces of P Granule components found in  membrane bound somatic cells Conclusion: P granule components are degraded by autophagy
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1.) How they form in somatic cells
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1B.) Gradual formation of P granules Methods:Immunostaining with K76 Results
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1B.) Gradual formation of P granules
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