Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Transcription profiling of mosquitos fed blood infected with two alternative P. berghei strains: wild type (wt) or an invasion-deficient, CTRP (Circumsporozoite- and TRAP-related protein) knockout (ko) strain

ABSTRACT: Female mosquitoes fed with blood infected with two alternative P. berghei strains: wild type (wt) or an invasion-deficient, CTRP (Circumsporozoite- and TRAP-related protein) knockout (ko) strain. We used CTRPko-infected midguts as reference, as they experience pre-invasion conditions similar to those of wt-infected midguts. Therefore, wt/CTRPko comparisons should reveal transcriptional changes specifically associated with parasite midgut invasion. We collected midguts at 18-22 h post infection (B, before midgut invasion), 24-28 h (I; during invasion) and 40-44 h (A; after invasion) and used them to hybridize two DNA microarrays: 4K microarrays containing 3,840 EST clones and MMC1 (or 20K) microarrays containing the same plus 15,840 additional ESTs. Two different biological experiments and two technical (dye-swap) replicates were performed for each array platform. In each experiment, the six different samples were used in seven different combinations to hybridize DNA microarrays, Thus, 3,840 ESTs that are represented in both array platforms were assessed in eight independent experimental replicates, whereas four replicates for the 15,840 ESTs that are unique to MMC1 were assessed in four independent experiments.

ORGANISM(S): Anopheles gambiae

DISEASE(S): Plasmodium berghei CTRP knockout strain (Dessens et al., 1999) infection

SUBMITTER: Fotis C. Kafatos

PROVIDER: E-MEXP-378 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Transcriptomic and functional analysis of an autolysis-deficient, teicoplanin-resistant derivative of methicillin-resistant Staphylococcus aureus.

Renzoni Adriana A Barras Christine C François Patrice P Charbonnier Yvan Y Huggler Elzbieta E Garzoni Christian C Kelley William L WL Majcherczyk Paul P Schrenzel Jacques J Lew Daniel P DP Vaudaux Pierre P

Antimicrobial agents and chemotherapy 20060901 9

The molecular basis of glycopeptide-intermediate S. aureus (GISA) isolates is not well defined though frequently involves phenotypes such as thickened cell walls and decreased autolysis. We have exploited an isogenic pair of teicoplanin-susceptible (strain MRGR3) and teicoplanin-resistant (strain 14-4) methicillin-resistant S. aureus strains for detailed transcriptomic profiling and analysis of altered autolytic properties. Strain 14-4 displayed markedly deficient Triton X-100-triggered autolysi ...[more]

PMID: 16940101

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Project description:Malaria is caused by Plasmodium parasites, which are transmitted via the bites of infected Anopheline mosquitoes. Midgut invasion is a major bottleneck for Plasmodium development inside the mosquito vectors as a rapidly responding immune system recognizes ookinetes and recruits killing factors from the midgut and surrounding tissues, dramatically reducing the population of invading ookinetes before they can successfully traverse the midgut epithelium. Understanding molecular details of the parasite-vector interactions requires precise measurement of nascent protein synthesis in the mosquito during Plasmodium infection. Current expression profiling primarily monitors alterations in steady-state levels of mRNA, but does not address the equally critical issue of whether the proteins encoded by the mRNAs are actually synthesized. In this study, we used sucrose density gradient centrifugation to isolate actively translating Anopheles gambiae mRNAs based upon their association with polyribosomes (polysomes). The proportion of individual gene transcripts associated with polysomes, which is determined by RNA deep sequencing, reflects mRNA translational status. This approach led to identification of 1017 mosquito transcripts that were primarily regulated at the translational level after ingestion of Plasmodium falciparum-infected blood. Caspar, a negative regulator of the NF-kappaB transcription factor Rel2, appears to be substantially activated at the translational levels during Plasmodium infection. In addition, transcripts of Dcr1, Dcr2 and Drosha, which are involved in small RNA biosynthesis, exhibited enhanced associations with polysomes after P. falciparum challenge. This observation suggests that mosquito microRNAs may play an important role in reactions against Plasmodium invasion. We analyzed both total cellular mRNAs and mRNAs that are associated with polysomes to simultaneously monitor transcriptomes and nascent protein synthesis in the mosquito. This approach provides more accurate information regarding the rate of protein synthesis, and identifies some mosquito factors that might have gone unrecognized because expression of these proteins is regulated mainly at the translational level rather than at the transcriptional level after mosquitoes ingest a Plasmodium-infected blood meal. Midguts from female Anopheles gambiae mosquitoes were dissected at around 24 h after ingestion of P. falciparum-infected blood. Mosquitoes fed on uninfected blood were used as control. A portion of the midguts were used for isolation of total cellular RNA. Extracts of the remaining midguts were fractionated over sucrose density gradients, and fifteen fractions were collected from the top of each gradient. Non-polysomal fractions, as well as polysomal fractions were combined, respectively, to obtain two RNA pools per gradient for three independent experiments. The first, last and the sample between non-polysomal and polysomal, were all discarded to ensure pure pools from each set. A fourth experiment was conducted where the polysomal and non-polysomal samples were not pooled, to be used later for qRT-PCR. The mRNA levels of individual mosquito genes in polysome (PS) fractions, nonpolysome (NP) fractions and unfractionated steady-state (total) RNA were determined using high throughput RNA deep sequencing. Each RNA pool generated 2.1-9.8 million raw read . Signal intensities of transcripts from each PS pool were compared to those of transcripts from a matching NP pool. To quantify the translational status of individual mRNA species, we define the relative PS loading [PL=P/(P+NP)] as the extent of mRNA association with polysomes. PL values were compared between the mosquitoes fed on P. falciparum-infected or -uninfected blood.

Dataset Information

Transcription profiling of mosquitos fed blood infected with two alternative P. berghei strains: wild type (wt) or an invasion-deficient, CTRP (Circumsporozoite- and TRAP-related protein) knockout (ko) strain

Publications

Transcriptomic and functional analysis of an autolysis-deficient, teicoplanin-resistant derivative of methicillin-resistant Staphylococcus aureus.

Similar Datasets

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