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. 2009 Mar 25;4(3):e4995. doi: 10.1371/journal.pone.0004995

Figure 1. Complexity of PCA3 transcripts.

Figure 1

(A) Partial PCA3 gene structure as originally reported by Bussemakers et al. [15] with 4 exons (open boxes ∼ not to scale) with alternate splicing of exon 2 and three alternate transcription termination sites in exon 4. 5′ RACE experiments (Supplementary Fig. S1) identified 4 novel PCA3 transcription initiation sites (isoforms 1–4 marked by vertical arrows pointing down with nucleotide sequence below located 1150 bp, 699 bp, 640 bp and 136 bp upstream of the original initiation site (renamed here isoform 5). 3′ RACE identified four novel polyadenylation sites (7 in total*) located in exon 4. The size of exon 1 is expanded from the original 120 bp to 1270 bp. Isoform 4 (PCA3-4) is the most highly expressed of the four novel isoforms. Four overlapping ORFs initiate from a single ‘ATG’ start site (vertical arrow pointing up) within PCA3-4 and terminate within one of the alternatively spliced exons (2a or 2b or 2c) or within exon 3. (B) RT-PCR amplification of BPH, PCa and PCa metastasis samples using a forward primer from within the novel PCA3-4 transcription start site and a reverse primer from novel exon 2a. (C) Complete structure of the PCA3 gene. Shading identifies the newly identified regions of the PCA3 gene which has 6 exons with alternate splicing of exon 2a (93 bp) and exon 2b (93 bp) and exon 2c (original exon 2, 165 bp). and (D) RT-PCR amplification of PCA3 using the same forward primer and a reverse primer from novel exon 2b and (E) RT-PCR amplification of PCA3 using a forward primer from PCA3-5F (within the original transcription start site) and a reverse primer spanning exons 1 and 3 [15].