Herpes simplex virus-induced early proteins
Başlık:
Herpes simplex virus-induced early proteins
Yazar:
MacDonald, Donald, author.
ISBN:
9780438059887
Yazar Ek Girişi:
Fiziksel Tanımlama:
1 electronic resource (212 pages)
Genel Not:
Source: Dissertation Abstracts International, Volume: 76-08C.
Advisors: Howard S. Marsden.
Özet:
Proteins induced early in infection by HSV have been studied. Under conditions designed to generate immediate-early (IE) polypeptides (Preston et al., 1978), eleven HSV-1 induced polypeptides were detected in BHK cells. Of these, five (IE 175, 136' (143), 110, 63 and 12) were usually induced in major amounts, whilst the other six (IE 68, 28, 25, 23, 16. 5 and 12. 7) were usually induced in lesser and more variable amounts. A markedly similar polypeptide profile was induced at the NPT by the HSV-1 ts mutant ts K. Four polypeptides (Vmw 175, 136' (143), 110 and 63) were overproduced continuously throughout infection and comigrated with four of the major IE polypeptides. In addition, polypeptides Vmw 68, 28, 25, 23, 16.5 and 12.7 also comigrated with IE polypeptides. The ts K polypeptide profile differed from that induced by ts+ virus under conditions designed to generate IE polypeptides in two main aspects: (1) Vmw 155, 117 and 73 were observed in BHK cells infected by ts K at 38° but not in BHK cells infected by ts+ virus under conditions designed to generate IE polypeptides, and (2) IE 12 was induced in BHK cells infected by ts+ virus under conditions designed to generate IE polypeptides but not in BHK cells infected by ts K at 38°. However, under conditions designed to generate IE polypeptides, ts K did induce a polypeptide which migrated slightly slower than the ts+ virus induced IE 12. IE 12 partitions with the cytoplasmic fraction of infected cells. Analysis of intertypic recombinants showed IE 12 mapped within the coordinates 0.824 - 0.874 and 0.930 - 0.990. The number and amounts of both IE polypeptides induced by ts+ virus and polypeptides induced by ts K at the NPT varied between experiments. The reasons for this variation were not clear but in ts K infections it was found not to depend upon small temperature deviations (+/- 1°C) from the NPT, and for IE polypeptides it was found not to depend upon the MOI. Pereira et al. (1977) observed the mobility of two pulse labelled cx polypeptides changed slightly after a 3 hr. chase. Similar experiments were performed in this study and in agreement with Pereira et al. (1977) three forms (a, b and c) of IE 175 (ICP4) and two forms (a and b) of IE 136' (betaICP6) were observed. In addition, two forms of IE 110 and IE 63 are shown in this Thesis. Mutants ts D, K and T induce polypeptides which comigrate with the a forms of IE 175, 136' (143), 110 and 63. After a 3 hr. chase at 38° IE 136' (143)a, 110a and 63 are fully processed to the more slowly migrating form. In contrast IE 175a induced by all three mutants is only partly processed and kept blocked in the intermediate b form. When ts K infected BHK cells were pulse-labelled with 35S-methionine and chased at 38° in unlabelled medium, a slow loss of label from the ts K-induced polypeptides Vmw 175, 136' (143), 110, 63 and 28 was observed and three new polypeptides, Vmw 100, 90 and 26 appeared. The three breakdown products accumulate almost exclusively in the nucleus. Tryptic peptide mapping showed that both Vmw100 and Vmw90 were derived from Vmw136' (143) and that Vmw26 was derived from Vmw63. Breakdown was observed in both exponentially growing and serum starved BHK cells. It could be inhibited by TPCK. The presence of metabolic inhibitors (actinomycin-D, cycloheximide or sodium azide) during the chase did not prevent the processing suggesting that the putative protease was already present at the start of the chase period. There was less breakdown of the IE polypeptides induced by ts+ virus than of the polypeptides induced by ts K at NPT. However, IE 110, 63, 28 and 12 were clearly unstable and IE 26 and IE 90 were accumulated during the chase.
Notlar:
School code: 0547
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Yer Numarası | Demirbaş Numarası | Shelf Location | Lokasyon / Statüsü / İade Tarihi |
---|---|---|---|
XX(684765.1) | 684765-1001 | Proquest E-Tez Koleksiyonu | Arıyor... |
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