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Investigation of phosphatidylinositol-3-kinase (PI3K) and extracellular signal-regulated kinase 1/2 (ERK1/2) activation Chiu, Doris Diane
Abstract
There have been conflicting studies in the past several years regarding the relationship between PI3K activation and its potential role in ERK1/2 activation. Some studies indicate that PI3K activation is required for ERK1/2 activation, yet other studies suggest that there is no relationship between the two. In this thesis we hypothesize that PI3K activation is not required for ERK1/2 activation. To investigate this hypothesis we used several cell systems, including a human, hemopoietic cell line, TF-1, and murine, hemopoietic cell lines FDC-P1 and BAF-3 cells. Also, we used a human embryonic kidney cell line, HEK 293. With the use of pharmacological inhibitors of PI3K, LY294002 and wortmannin it was concluded that ERK1/2 activation does not require PI3K activation. We also addressed the question of what role PI3K plays in the regulation of protein synthesis. However, in order to test this, we first investigated whether our cell systems contained the important components of the translational protein machinery including, elF-4E and 4E-BPs. Afterwards, we tested the effects of using various inhibitors, including the PI3K inhibitors LY294002, wortmannin, MEK1/2 inhibitors, U0I26 and the FRAP/mTOR inhibitor rapamycin. From our preliminary experiments we determined that 4E-BP1 is present in TF-1 cells and 4E-BP2 is present in FDC-P1, MC-9 and TF-1 cells. Furthermore, elF-4E is present in all three cell systems. With the use of the inhibitors, we observed that LY294002 inhibits the phosphorylation of 4E-BP1, U0126 did not block the phosphorylation of 4E-BP1 and rapamycin partially blocked the phosphorylation of 4E-BP1. Surprisingly, wortmannin did not block the phosphorylation of 4EBP1. Due to the preliminary nature of the results, more experiments are needed to fully understand the role that PI3K may play in the regulation of protein translation.
Item Metadata
Title |
Investigation of phosphatidylinositol-3-kinase (PI3K) and extracellular signal-regulated kinase 1/2 (ERK1/2) activation
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Creator | |
Publisher |
University of British Columbia
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Date Issued |
2001
|
Description |
There have been conflicting studies in the past several years regarding
the relationship between PI3K activation and its potential role in ERK1/2
activation. Some studies indicate that PI3K activation is required for ERK1/2
activation, yet other studies suggest that there is no relationship between the
two. In this thesis we hypothesize that PI3K activation is not required for ERK1/2
activation. To investigate this hypothesis we used several cell systems, including
a human, hemopoietic cell line, TF-1, and murine, hemopoietic cell lines FDC-P1
and BAF-3 cells. Also, we used a human embryonic kidney cell line, HEK 293.
With the use of pharmacological inhibitors of PI3K, LY294002 and wortmannin it
was concluded that ERK1/2 activation does not require PI3K activation.
We also addressed the question of what role PI3K plays in the regulation
of protein synthesis. However, in order to test this, we first investigated whether
our cell systems contained the important components of the translational protein
machinery including, elF-4E and 4E-BPs. Afterwards, we tested the effects of
using various inhibitors, including the PI3K inhibitors LY294002, wortmannin,
MEK1/2 inhibitors, U0I26 and the FRAP/mTOR inhibitor rapamycin. From our
preliminary experiments we determined that 4E-BP1 is present in TF-1 cells and
4E-BP2 is present in FDC-P1, MC-9 and TF-1 cells. Furthermore, elF-4E is
present in all three cell systems. With the use of the inhibitors, we observed that
LY294002 inhibits the phosphorylation of 4E-BP1, U0126 did not block the
phosphorylation of 4E-BP1 and rapamycin partially blocked the phosphorylation
of 4E-BP1. Surprisingly, wortmannin did not block the phosphorylation of 4EBP1.
Due to the preliminary nature of the results, more experiments are needed
to fully understand the role that PI3K may play in the regulation of protein
translation.
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Extent |
5608031 bytes
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Genre | |
Type | |
File Format |
application/pdf
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Language |
eng
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Date Available |
2009-08-04
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Provider |
Vancouver : University of British Columbia Library
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Rights |
For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https://open.library.ubc.ca/terms_of_use.
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DOI |
10.14288/1.0090096
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URI | |
Degree | |
Program | |
Affiliation | |
Degree Grantor |
University of British Columbia
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Graduation Date |
2001-11
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Campus | |
Scholarly Level |
Graduate
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Aggregated Source Repository |
DSpace
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Item Media
Item Citations and Data
Rights
For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https://open.library.ubc.ca/terms_of_use.