Figure 5
shows that wild type and mutant
E1A proteins localized to the nucleus in all of the constructs examined,
consistent with the presence in each of a C-terminal nuclear localization site.
Figure 5
Cellular localization of E1A proteins.
Growing cultures of MSY596 were transferred to media containing 2% galactose
for four hours to express E1A proteins,
and then plated on glass cover slips.
Identical fields were analyzed by phase contrast microscopy (Cells),
indirect immunofluorescence using the anti-E1A monoclonal antibody M73
and FITC conjugated anti-IgG as stain (E1A),
and fluorescence microscopy using
4',6-Diamidino-2-phenylindole-dihydrochloride as stain (DNA).
(A)
E1A289;
(B)
E1A243;
(C)
dl1101;
(D)
dl1103;
(E)
dl1104
Cells expressing E1A243 and
dl1101
also showed increased cytoplasmic staining relative to the other derivatives;
increased cytoplasmic staining with E1A243 has been noted previously
(73).
These results show that
the loss of function seen with E1A243,
and with
dl1101,
dl1103,
and
dl1104,
can not be due to an absence of nuclear E1A protein.
Therefore,
we conclude that the inhibition of growth in G1
caused by expression of E1A proteins in
S. cerevisiae
depends on the N-terminal region,
CR1,
and CR3.
Next:Residues 1-82 are sufficient for function
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November 26, 1995 at 9:52 PM