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Cellular aspects of retroviral buddi...

Sherer, Nathan Mark.

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Cellular aspects of retroviral budding and transmission revealed by live cell imaging.

紀錄類型:	書目-電子資源 : Monograph/item
正題名/作者:	Cellular aspects of retroviral budding and transmission revealed by live cell imaging./
作者:	Sherer, Nathan Mark.
面頁冊數:	168 p.
附註:	Source: Dissertation Abstracts International, Volume: 67-04, Section: B, page: 1814.
Contained By:	Dissertation Abstracts International67-04B.
標題:	Biology, Cell. -
電子資源:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3214300
ISBN:	9780542653742

Cellular aspects of retroviral budding and transmission revealed by live cell imaging.
Sherer, Nathan Mark.

Cellular aspects of retroviral budding and transmission revealed by live cell imaging. - 168 p.

Source: Dissertation Abstracts International, Volume: 67-04, Section: B, page: 1814.

Thesis (Ph.D.)--Yale University, 2006.

The assembly and budding of retroviruses such as the murine leukemia virus (MLV) and human immunodeficiency virus-1 (HIV-1) is driven by oligomerization of the viral Gag polyprotein at the cytosolic face of cellular membranes. However, Gag must also recruit cellular factors linked to the vacuolar protein sorting machinery (vps) for efficient virus-cell fission during late stage budding. In these studies, live confocal imaging in conjunction with electron microscopy revealed that, in addition to the plasma membrane, MLV and HIV-1 Gag can undergo assembly intracellularly into the interior of late endosomal multivesicular bodies (MVBs), indicating that viruses can interact with the vps machinery in a traditional sense, as ubiquitinated cargo destined for sorting into endosomes. Because several modified MVBs such as MHC class II compartments are secretory in nature, aspects of MVB trafficking and biogenesis were tested in order to determine if MVB budding contributes productively to virus egress. MLV release was not affected by stimuli or factors related to Ca2+-regulated lysosomal exocytosis. While cytoplasmic acidification triggered the dramatic appearance of Gag-GFP particles at the cell surface, these particles were not released into the culture supernatant. In contrast, the targeting of the MVB-resident tetraspanin CD63 to the plasma membrane resulted in a two-fold increase in virus release, suggesting that MLV production may be favored by the retention of endosome-related platforms at the cell surface.

ISBN: 9780542653742Subjects--Topical Terms:

1017686
Biology, Cell.

Cellular aspects of retroviral budding and transmission revealed by live cell imaging.
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520 $a The assembly and budding of retroviruses such as the murine leukemia virus (MLV) and human immunodeficiency virus-1 (HIV-1) is driven by oligomerization of the viral Gag polyprotein at the cytosolic face of cellular membranes. However, Gag must also recruit cellular factors linked to the vacuolar protein sorting machinery (vps) for efficient virus-cell fission during late stage budding. In these studies, live confocal imaging in conjunction with electron microscopy revealed that, in addition to the plasma membrane, MLV and HIV-1 Gag can undergo assembly intracellularly into the interior of late endosomal multivesicular bodies (MVBs), indicating that viruses can interact with the vps machinery in a traditional sense, as ubiquitinated cargo destined for sorting into endosomes. Because several modified MVBs such as MHC class II compartments are secretory in nature, aspects of MVB trafficking and biogenesis were tested in order to determine if MVB budding contributes productively to virus egress. MLV release was not affected by stimuli or factors related to Ca2+-regulated lysosomal exocytosis. While cytoplasmic acidification triggered the dramatic appearance of Gag-GFP particles at the cell surface, these particles were not released into the culture supernatant. In contrast, the targeting of the MVB-resident tetraspanin CD63 to the plasma membrane resulted in a two-fold increase in virus release, suggesting that MLV production may be favored by the retention of endosome-related platforms at the cell surface.
520 $a Additionally, time-lapse imaging of fluorescent MLV particles at the surface of infected cells revealed that viruses frequently retained low affinity associations with the plasma membrane after budding. Surprisingly, while the release of these viruses into the culture supernatant was rarely detected, viruses underwent efficient release and cell-to-cell transmission after binding to the peripheral filopodia of neighboring uninfected target cells. This mode of transmission required high affinity interactions between the viral Envelope glycoprotein and receptor on the target cell, triggering strong actin-dependent retrograde pulling forces. Upon release, viruses surfed rapidly and directionally towards the cell body of uninfected cells. As such, we provide evidence for a new model of cell-to-cell virus transmission regulated by relative virus-cell affinities and driven by peripheral retrograde actin flow.
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