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Referee comments: Referee 1

Posted by PLOS_ONE_Group on 15 Feb 2008 at 18:18 GMT

Referee 1's review:

The manuscript by Pezer and Ugarkovic entitled "RNA Pol II promotes transcription of centromeric satellite DNA in beetles" reports the observation that centromeric satellite DNA (PRAT) is transcribed in this organism. PRAT is expressed at similar levels development, however, the forward transcript is ~10X more abundant than the reverse. Interestingly, the RNAi pathway does not process the transcripts. Multiple transcription initiation sites and polyadenylation signals have been identified, and transcription is sensitive to an inhibitor of RNA Pol II transcription, a-amanitin. The sequencing of transcription products, and assembly of a phylogenetic tree, revealed no transcription bias among the satellite subfamilies.

These findings are clearly presented and placed within the context of current centromeric satellite literature.

Points of concern:

1. Overall, the figures are very small and difficult to read. In particular, Figures 5B and 6B, which are mostly text, need to be enlarged or otherwise improved before publication.

2. The authors demonstrate that only ~20% of PRAT transcripts are polyadenylated, suggesting that RNA pol I or pol III may also act upon PRAT DNA. The authors should examine the PRAT sequence for the presence of A and B box consensus sites, which are associated with RNA pol III transcription. It should also be noted that both RNA pol III association and Pol III transcripts (tRNA genes) have been identified at the S. pombe centromere (Noma et al., 2006; Scott et al., 2006).

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N.B. These are the comments made by the referee when reviewing an earlier version of this paper. Prior to publication the manuscript has been revised in light of these comments and to address other editorial requirements.