Poly A- transcripts expressed in Hela cells

Qingfa Wu, Yeong C. Kim, Jian Lu, Zhenyu Xuan, Jun Chen, Yonglan Zheng, Tom Zhou, Michael Q. Zhang, Chung I. Wu, San Ming Wang

Research output: Contribution to journalArticle

69 Scopus citations

Abstract

Background: Transcripts expressed in eukaryotes are dissified as poly A+ transcripts or poly A- transcripts based on the presence or absence f the 3′ poly A tail. Most transcripts identified so far are poly A+ transcripts, whereas the poly A- transcripts remain largely unknown. Methodology/Principal Findings: We developed the TRD (Total RNA Detection) system for transcript identification. The system detects the transcripts through the following steps: 1) depleting the abundant ribosomal and small-size transcripts; 2) synthesizing cDNA without regard to the status of the 3′ poly A tail; 3) applying the 454 sequencing technology for massive 3′ EST collection from the cDNA; and 4) determining the genome origins of the detected transcripts by mapping the sequences to the human genome reference sequences. Using this system, we characterized the cytoplasmic transcripts from HeLa cells. Of the 13,467 distinct 3′ ESTs analyzed, 24% are poly A-, 36% are poly A+, and 40% are bimorphic with poly A+ features but without the 3′ poly A tail. Most of the poly A- 3′ ESTs do not match known transcript sequences; they have a similar distribution pattern in the genome as the poly A+ and bimorphic 3′ ESTs, and their mapped intergenic regions are evolutionarily conserved. Experiments confirmed the authenticity of the detected poly A- transcripts. Conclusion/Significance: Our study provides the first large-scale sequence evidence for the presence of poly A- transcripts in eukaryotes. The abundance of the poly A- transcripts highlights the need for comprehensive identification of these transcripts for decoding the transcriptome, annotating the genome and studying biological relevance of the poly A- transcripts.

Original languageEnglish (US)
Article numbere2803
JournalPloS one
Volume3
Issue number7
DOIs
StatePublished - Jul 30 2008
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

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    Wu, Q., Kim, Y. C., Lu, J., Xuan, Z., Chen, J., Zheng, Y., Zhou, T., Zhang, M. Q., Wu, C. I., & Wang, S. M. (2008). Poly A- transcripts expressed in Hela cells. PloS one, 3(7), [e2803]. https://doi.org/10.1371/journal.pone.0002803