α-Actinin TvACTN3 of Trichomonas vaginalis Is an RNA-Binding Protein That Could Participate in Its Posttranscriptional Iron Regulatory Mechanism
Figure 4
Effects of iron on the transcription of T. vaginalis actinin genes. (a) RT-PCR with specific primers for each T. vaginalis actinin gene (tvactn1, tvactn2, tvactn3, tvactn4, and tvactn5; Table 1) using cDNA from parasites grown under iron-depleted (0 μM; lane 1), normal (20 μM; lane 2), and iron-rich (250 μM; lane 3) conditions (tvactn1 to tvactn5). RT-PCR with specific primers for the β-tubulin gene using the same cDNA as an internal control. tvpfo a and tvcp12 genes were used as control genes that are overexpressed under iron-rich or iron-depleted conditions, respectively. A reverse transcriptase minus [RT-PCR (−)] reaction was used to verify the lack of gDNA contamination in the cDNA samples using the same pairs of primers for each gene; gDNA was used as a positive control (+), except in RT-PCR (−), in which no DNA was added. PCRs without gDNA were used as negative controls (−). The sizes of the amplicons are given in bp. (b) qRT-PCR was used to quantify the different levels of the five tvactn mRNAs in trichomonads grown under different iron conditions. Bars represent the standard error of triplicated samples. Asterisks (*) or (**) compared iron-depleted with iron-rich or normal conditions.
