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Series GSE7223

Query DataSets for GSE7223

Status

Public on Oct 31, 2007

Title

Genes regulated by the processed form of AIbZIP/CREB3L4 in LNCaP prostate cancer cells

Organism

Homo sapiens

Experiment type

Expression profiling by array

Summary

Androgen-induced bZIP (AIbZIP) is a basic leucine zipper (bZIP)
transcription factor that is encoded by the CREB3L4 (cAMP responsive element
binding protein 3-like 4) gene. In humans, AIbZIP mRNA is most abundant in
the prostate but it is also present, albeit at much lower levels, in the
pancreas and other tissues. The full-length AIbZIP protein localizes to the
endoplasmic reticulum and the available data suggest that AIbZIP is
processed to its transcriptionally active form by regulated intramembrane
proteolysis (RIP). To define the transcriptional programme that is initiated
by AIbZIP, we generated LNCaP prostate cancer cell lines that conditionally
express the processed form of AIbZIP. The recombinant protein consists of
the first 290 amino acids of AIbZIP fused to a C-terminal HA epitope.
Conditional expression was achieved using the RheoSwitch expression system.
Clone number 7-11 was exposed to the inducing agent (RSL1) for 6 and 24 hr
and Affymetrix microarrays were used to identify AIbZIP-regulated
transcripts. Candidate genes were identified by comparing the probe set
intensities of RSL1-treated cells and control cells (cells exposed to
vehicle). Forty-eight genes were then validated by Northern blot analysis.
Keywords: time course

Overall design

Clone 7-11 cells (passage 42) were plated in 24 10-cm culture dishes.
Sixteen dishes were cultured in medium containing RSL1 (500 nM) and eight
dishes were cultured in medium containing an equivalent amount of the RSL1
vehicle (DMSO). Eight RSL1-treated dishes were harvested 6 hr after addition
of RSL1 and another eight dishes were harvested 24 hr after addition of
RSL1. At each time point, the dishes were pooled two by two to yield a total
of 12 individual samples i.e. 4 controls, 4 exposed to RSL1 for 6 hr, and 4
exposed to RSL1 for 24 hr. One sample of each group was processed to nuclear
extracts to assess the levels of recombinant AIbZIP by immunoblotting. The
three other samples of each group were processed separately to cRNA probes
which were then hybridized to Affymetrix Human Genome U133 Plus 2.0 arrays
(Batch nos. 4003850 and 4003847).

Web link

http://www.crchul.ulaval.ca/

Contributor(s)

Labrie C, Calvo E

Citation(s)

17712038

Submission date

Mar 08, 2007

Last update date

Mar 25, 2019

Contact name

Ezequiel L Calvo