Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Transcription profiling of cat FIV infected lymphocytes at 1, 2, 4, and 24hrs post infection

ABSTRACT: Gene Expression Changes in Lymphocytes at Various Times Post-Feline Immunodeficiency Virus Infection:

Lymphocytes were cultured for 3 to 5 days with hrIL-2 and Con-A, counted, and prepared at a density of 4.0 x 10e6 cells/ml of media. Approximately 10e7 cells were seeded into eight separate 50 ml centrifuge tubes, and cells were treated with polybrene for 1 hour at 37oC.
The cells were washed and infected with 16 TCID50 of USgaB01 for 2 hours, or mock infected with "spent" media. The cells were washed, re-suspended in 10 ml of fresh media, and aliquots were cultured for 1, 2, 4, and 24 hours. Total RNA was isolated at each time
point for microarray analysis. Individual aliquots of lymphocytes were infected on 3 separate
occasions.

ORGANISM(S): Felis catus

SUBMITTER: Ryan J Dowling

PROVIDER: E-MEXP-204 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Gene-expression changes induced by Feline immunodeficiency virus infection differ in epithelial cells and lymphocytes.

Dowling R J O RJO Bienzle D D

The Journal of general virology 20050801 Pt 8

Infection of cats with Feline immunodeficiency virus (FIV) is an important model for understanding comparative lentivirus biology. In vivo, FIV infects lymphocytes and monocyte/macrophages, but in vitro infection is commonly investigated in epithelial Crandell-Reese Feline Kidney (CRFK) cells. In this study, the transcriptional responses of CRFK cells and primary lymphocytes to infection with FIV 34TF, a cloned subtype A virus, and FIV USgaB01, a biological subtype B isolate, were determined. Re ...[more]

PMID: 16033971

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Project description:Copy number expansions such as amplifications and duplications contribute to human phenotypic variation, promote molecular diversification during evolution, and drive the initiation and/or progression of various cancers. The mechanisms underlying these copy number changes are still incompletely understood, however. We recently demonstrated that transient, limited re-replication from a single origin in Saccharomyces cerevisiae efficiently induces segmental amplification of the re-replicated region. Structural analyses of such re-replication induced gene amplifications (RRIGA) suggested that RRIGA could provide a new mechanism for generating copy number variation by non-allelic homologous recombination (NAHR). Here we elucidate this new mechanism and provide insight into why it is so efficient. We establish that sequence homology is both necessary and sufficient for repetitive elements to participate in RRIGA and show that their recombination occurs by a single-strand annealing (SSA) mechanism. We also find that re-replication forks are prone to breakage, accounting for the widespread DNA damage associated with deregulation of replication proteins. These breaks appear to stimulate NAHR between re-replicated repeat sequences flanking a re-initiating replication origin. Our results support a RRIGA model where the expansion of a re-replication bubble beyond flanking homologous sequences followed by breakage at both forks in trans provides an ideal structural context for SSA–mediated NAHR to form a head-to-tail duplication. Given the remarkable efficiency of RRIGA, we suggest it may be an unappreciated contributor to copy number expansions in both disease and evolution. The arrays in this series are primary comparative genomic hybridizations to determine genomic changes after re-replication. Genomic DNA was purified from reference Saccharomyces cerevisiae cells or survivors of re-replication, differentially labeled with Cy3 and Cy5, and competitively hybridized to a spotted microarray containing ORF and intergenic PCR products. Cy5/Cy3 ratios (or the inverse in the case of dye swapped samples) are normalized so that the average ratio of all elements was 1 (except for arrays measuring re-replication induction where this ratio was set to 2). A small number of the arrays were used to determine the extent and location of re-replication under different conditions. For those, genomic DNA was purified from non-replicating and re-replicating cells and treated as above. Series contains a total of 333 hybridizations. Note that the VALUE field reported for all of the sample tables in this series are the log2 of the normalized Cy5/Cy3 (or Cy3/Cy5) ratio. To convert these into usable copy number profiles raise 2 to the indicated VALUE.

Dataset Information

Transcription profiling of cat FIV infected lymphocytes at 1, 2, 4, and 24hrs post infection

Publications

Gene-expression changes induced by Feline immunodeficiency virus infection differ in epithelial cells and lymphocytes.

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