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Series GSE2296 Query DataSets for GSE2296
Status Public on Feb 10, 2006
Title Effects of inducible HIV-1 Vpr protein on cellular gene expression
Organism Homo sapiens
Experiment type Expression profiling by array
Summary Three 293 cell-derivative cell line were constructed using commercially available Invitrogen Flp-In TREx 293 cells. The cell lines express wild type HIV-1 Vpr and two mutant Vprs, F72A/R73A and R80A. The mutant Vprs are defective in the Vpr cell cycle arrest phenotype.
The cell lines were characterized for their ability to express Vpr at the RNA and protein levels and functionally for the ability of the inducibly expressed Vprs to arrest cell cycle progression at G2/M. To study whether certain host cell genes showed alterations in expression following induced expression of Vpr, we conducted a large scale host cell gene expression profiling study using microarray technologies. To determine whether the differential expression of certain genes was associated with Vpr-induced cell cycle arrest, we compared the gene expression pattern of wild type Vpr expressing cells with the pattern of Vpr mutant F72A/R73A- or R80A-Vpr expressing cells. Doxycycline-induced and uninduced and Flp-In TREx 293/Vpr cells were collected 0, 1, 2, 4, 6, 8, 12, 16 and 24 hours post induction (hpi). Doxycyclineinduced and uninduced Flp-In TREx 293/Blank cells were also collected as a reference for following experiments. Total RNA was isolated using RNAeasy Midi Kit (QIAGEN), according to the manufacturer's protocol. Human long oligonucleotide microarrays were obtained from the National Cancer Institute Microarray Facility, Advanced Technology Center (Gaithersburg, MD). The microarrays (Hs Operon V2) contained 22,434 oligonucleotide (60-70 mer) spots on a glass slide. 25 µg of total RNAs isolated from doxycycline-induced Flp-In TREx 293 stably transfected blank- and FLAG tagged-Vpr cells were reverse transcribed using an oligo (dT) primer and aminoallyl-dNTP (Amersham Pharmacia). Unincorporated dNTPs were removed with QIAquick PCR purification kit (QIAGEN).
Purified cDNA was processed for coupling reaction with Cy 5 or Cy 3-ester dye, respectively.
Keywords: other
 
 
Contributor(s) Yoshizuka N
Citation(s) 16103188
Submission date Feb 18, 2005
Last update date Jan 17, 2013
Contact name Naoto Yoshizuka
E-mail(s) yoshizun@mail.nih.gov
Phone 301-402-3624
Organization name NCI
Street address
City Bethesda
State/province MD
ZIP/Postal code 20892
Country USA
 
Platforms (1)
GPL1528 NCI/ATC Hs-OperonV2
Samples (87)
GSM41859 R80A_0h:Blank_0h - mAdb ID 55193
GSM41860 R80A_1h:Blank_1h - mAdb ID 55194
GSM41861 R80A_2h:Blank_2h - mAdb ID 55195
Relations
BioProject PRJNA91673

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