Comparative expression analysis uncovers novel features of endogenous antisense transcription.

Increasing numbers of sense-antisense transcripts (SATs), which are transcribed from the same chromosomal location but in opposite directions, have been identified in various eukaryotic species, but the biological meanings of most SATs remain unclear. To improve understanding of natural sense-antisense transcription, we performed comparative expression profiling of SATs conserved among humans and mice. Using custom oligo-arrays loaded with probes that represented SATs with both protein-coding and non-protein-coding transcripts, we showed that 33% of the 291 conserved SATs displayed identical expression patterns in the two species. Among these SATs, expressional balance inversion of sense-antisense genes was mostly observed in testis at a tissue-specific manner. Northern analyses of the individual conserved SAT loci revealed that: (i) a smeary hybridization pattern was present in mice, but not in humans, and (2) small RNAs (about 60 to 80 nt) were detected from the exon-overlapping regions of SAT loci. In addition, further analyses showed marked alteration of sense-antisense expression balance throughout spermatogenesis in testis. These results suggest that conserved SAT loci are rich in potential regulatory roles that will help us understand this new class of transcripts underlying the mammalian genome.

[1]  Shoshi Kikuchi,et al.  Antisense transcripts with rice full-length cDNAs , 2003, Genome Biology.

[2]  M. Nóbrega,et al.  Comparative genomics at the vertebrate extremes , 2004, Nature Reviews Genetics.

[3]  Mouse Genome Sequencing Consortium Initial sequencing and comparative analysis of the mouse genome , 2002, Nature.

[4]  P. Stadler,et al.  RNA Maps Reveal New RNA Classes and a Possible Function for Pervasive Transcription , 2007, Science.

[5]  T. Hughes,et al.  A systematic search for new mammalian noncoding RNAs indicates little conserved intergenic transcription , 2005, BMC Genomics.

[6]  D. Botstein,et al.  Cluster analysis and display of genome-wide expression patterns. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[7]  Oliver H. Tam,et al.  Characterization of Dicer-deficient murine embryonic stem cells. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[8]  D. Higgs,et al.  Transcription of antisense RNA leading to gene silencing and methylation as a novel cause of human genetic disease , 2003, Nature Genetics.

[9]  Erez Y. Levanon,et al.  Widespread occurrence of antisense transcription in the human genome , 2003, Nature Biotechnology.

[10]  H. Kiyosawa,et al.  Speculations on the role of natural antisense transcripts in mammalian X chromosome evolution , 2003, Cytogenetic and Genome Research.

[11]  Michal Galdzicki,et al.  Mammalian overlapping genes: the comparative perspective. , 2004, Genome research.

[12]  Michael Ashburner,et al.  Annotation of the Drosophila melanogaster euchromatic genome: a systematic review , 2002, Genome Biology.

[13]  M. Lazar,et al.  Post-transcriptional Regulation of Thyroid Hormone Receptor Expression by cis-Acting Sequences and a Naturally Occurring Antisense RNA* , 2000, The Journal of Biological Chemistry.

[14]  L. Romrell,et al.  Separation of mouse spermatogenic cells by sedimentation velocity. A morphological characterization. , 1976, Developmental biology.

[15]  Sin Lam Tan,et al.  Complex Loci in Human and Mouse Genomes , 2006, PLoS genetics.

[16]  O. Borsani,et al.  Endogenous siRNAs Derived from a Pair of Natural cis-Antisense Transcripts Regulate Salt Tolerance in Arabidopsis , 2005, Cell.

[17]  S. Batalov,et al.  Antisense Transcription in the Mammalian Transcriptome , 2005, Science.

[18]  Joseph M. Dale,et al.  Empirical Analysis of Transcriptional Activity in the Arabidopsis Genome , 2003, Science.

[19]  Satoshi Tanaka,et al.  Non-coding RNA directed DNA demethylation of Sphk1 CpG island. , 2004, Biochemical and biophysical research communications.

[20]  P. Glenister,et al.  Identification of an imprinting control region affecting the expression of all transcripts in the Gnas cluster , 2006, Nature Genetics.

[21]  S. Brantl,et al.  Antisense-RNA regulation and RNA interference. , 2002, Biochimica et biophysica acta.

[22]  Ben Lehner,et al.  In search of antisense. , 2004, Trends in biochemical sciences.

[23]  M. Tomita,et al.  Comparative analysis of cis-encoded antisense RNAs in eukaryotes. , 2007, Gene.

[24]  Jun Kawai,et al.  Clusters of Internally Primed Transcripts Reveal Novel Long Noncoding RNAs , 2006, PLoS genetics.

[25]  Terrence S. Furey,et al.  The UCSC Genome Browser Database , 2003, Nucleic Acids Res..

[26]  Xiaoqiu Huang,et al.  Over 20% of human transcripts might form sense-antisense pairs. , 2004, Nucleic acids research.

[27]  G. Rubin,et al.  A computer program for aligning a cDNA sequence with a genomic DNA sequence. , 1998, Genome research.

[28]  K. Nishikura,et al.  Extensive adenosine‐to‐inosine editing detected in Alu repeats of antisense RNAs reveals scarcity of sense–antisense duplex formation , 2006, FEBS letters.

[29]  William Stafford Noble,et al.  Identification and analysis of functional elements in 1% of the human genome by the ENCODE pilot project , 2007, Nature.

[30]  Alok J. Saldanha,et al.  Java Treeview - extensible visualization of microarray data , 2004, Bioinform..

[31]  Songtao Jia,et al.  RNAi-Mediated Targeting of Heterochromatin by the RITS Complex , 2004, Science.

[32]  W. J. Kent,et al.  BLAT--the BLAST-like alignment tool. , 2002, Genome research.

[33]  M. Dym,et al.  Spermatogenic cells of the prepuberal mouse: isolation and morphological characterization , 1977, The Journal of cell biology.

[34]  Ronald H. A. Plasterk,et al.  Transposon silencing in the Caenorhabditis elegans germ line by natural RNAi , 2003, Nature.

[35]  Yitzhak Pilpel,et al.  Genome‐wide natural antisense transcription: coupling its regulation to its different regulatory mechanisms , 2006, EMBO reports.

[36]  Yoshihide Hayashizaki,et al.  Antisense transcripts with FANTOM2 clone set and their implications for gene regulation. , 2003, Genome research.

[37]  Miao Sun,et al.  Evidence for variation in abundance of antisense transcripts between multicellular animals but no relationship between antisense transcriptionand organismic complexity. , 2006, Genome research.

[38]  E. Birney,et al.  Analysis of the mouse transcriptome based on functional annotation of 60,770 full-length cDNAs , 2002, Nature.

[39]  Jeannie T. Lee,et al.  Tsix, a gene antisense to Xist at the X-inactivation centre , 1999, Nature Genetics.

[40]  R. Sorek,et al.  Is there any sense in antisense editing? , 2005, Trends in genetics : TIG.

[41]  Wolfgang Huber,et al.  A high-resolution map of transcription in the yeast genome. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[42]  J. Vaughn,et al.  RNA editing and regulation of Drosophila 4f-rnp expression by sas-10 antisense readthrough mRNA transcripts. , 2003, RNA.

[43]  Yoshihide Hayashizaki,et al.  Disclosing hidden transcripts: mouse natural sense-antisense transcripts tend to be poly(A) negative and nuclear localized. , 2005, Genome research.

[44]  Terry Gaasterland,et al.  Genome-wide prediction and identification of cis-natural antisense transcripts in Arabidopsis thaliana , 2005, Genome Biology.