Francesca Bottanelli

Institute:Department of Cell Biology, Yale University School of Medicine, New Haven USA
Title:Live-cell nanoscopy of protein sorting at the Golgi
Location:H.R.Kruytgebouw O622
Abstract:["There is a lack of understanding of the dynamics and nanoscale organization of the Golgi apparatus in living cells. We have recently developed a novel labeling strategy for dual-color live-cell stimulated emission depletion (STED) super-resolution imaging. Here we report that SNAP and Halo substrates of the dye ATTO590 can cross biological membranes to provide a second color needed to complement the well known live-cell STED-compatible dye silicone rhodamine (SiR). \r\nTaking advantage of gene editing techniques (CRISPR\/Cas9) to avoid over-expression and our novel live-cell STED labeling strategy we revisit the role of ARF1. Beside its well-established role in generating COPI vesicles by recruiting Coatomer at the Golgi, we find that ARF1 is additionally involved in the formation of anterograde and retrograde tubular carriers. The ARF1-positive tubular carriers can be divided into two subclasses: i) anterograde tubules that are also positive for Clathrin and contain the cargo VSV G, and ii) retrograde tubules that are positive for Coatomer and contain the cargo KDEL receptor. ARF1 tubular transport intermediates account for a major membrane flow out of the Golgi."]
Host:Bernd Helms - (0)30 253 5375