Identification of putative noncoding polyadenylated transcripts in Drosophila melanogaster.

Analysis of EST and cDNA collections from a number of metazoan species has identified genes encoding long polyadenylated transcripts that do not contain ORFs of lengths typical for protein-encoding mRNAs. Noncoding functions of such polyadenylated transcripts have been elucidated in only a few examples. The corresponding genes neither contain hallmark sequence motifs nor appear to have been conserved across phyla. Thus, it is impossible to systematically identify new members of this class of gene by using sequence homology and traditional gene-finding algorithms that depend on protein-coding potential. Consequently, even their approximate number has not been established for any metazoan genome. We curated polyadenylated transcripts with limited protein-coding capacity from intergenic regions of the Drosophila melanogaster genome. We used RT-PCR assays, hybridization to RNA blots and whole-mount embryos, and computational analyses to characterize candidate transcripts. We verify the structures and expression of 17 distinct, likely non-protein-coding polyadenylated transcripts. We show that the expression of many of these transcripts is conserved in other Drosophila species, indicating that they have important biological functions.

[1]  Inna Dubchak,et al.  Comparative genome sequencing of Drosophila pseudoobscura: chromosomal, gene, and cis-element evolution. , 2005, Genome research.

[2]  J. Clayton,et al.  The clock gene period of the housefly, Musca domestica, rescues behavioral rhythmicity in Drosophila melanogaster. Evidence for intermolecular coevolution? , 2000, Genetics.

[3]  Ziheng Yang,et al.  Phylogenetic Analysis by Maximum Likelihood (PAML) , 2002 .

[4]  P. Lasko,et al.  Requirement for a Noncoding RNA in Drosophila Polar Granules for Germ Cell Establishment , 1996, Science.

[5]  R. Kelley Path to equality strewn with roX. , 2004, Developmental biology.

[6]  E. Myers,et al.  Finishing a whole-genome shotgun: Release 3 of the Drosophila melanogaster euchromatic genome sequence , 2002, Genome Biology.

[7]  D. Hogness,et al.  Novel transcripts from the Ultrabithorax domain of the bithorax complex. , 1987, Genes & development.

[8]  Scott A. Rifkin,et al.  A Gene Expression Map for the Euchromatic Genome of Drosophila melanogaster , 2004, Science.

[9]  Bradley P. Coe,et al.  A tiling resolution DNA microarray with complete coverage of the human genome , 2004, Nature Genetics.

[10]  M. Tomita,et al.  Identification of putative noncoding RNAs among the RIKEN mouse full-length cDNA collection. , 2003, Genome research.

[11]  S. P. Fodor,et al.  Large-Scale Transcriptional Activity in Chromosomes 21 and 22 , 2002, Science.

[12]  B. Haas,et al.  Full-length messenger RNA sequences greatly improve genome annotation , 2002, Genome Biology.

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

[14]  Kanako O. Koyanagi,et al.  Integrative Annotation of 21,037 Human Genes Validated by Full-Length cDNA Clones , 2004, PLoS Biology.

[15]  Maciej Szymanski,et al.  Non-coding, mRNA-like RNAs database Y2K , 2000, Nucleic Acids Res..

[16]  G. Rubin,et al.  Computational identification of Drosophila microRNA genes , 2003, Genome Biology.

[17]  Sanghyuk Lee,et al.  MicroRNA genes are transcribed by RNA polymerase II , 2004, The EMBO journal.

[18]  S. Eddy,et al.  Computational identification of noncoding RNAs in E. coli by comparative genomics , 2001, Current Biology.

[19]  Jian Wang,et al.  The Genome Sequence of the Malaria Mosquito Anopheles gambiae , 2002, Science.

[20]  G. Rubin,et al.  The Drosophila melanogaster genome. , 2003, Annual review of genomics and human genetics.

[21]  C. Wilkerson,et al.  Identification and analysis of Arabidopsis expressed sequence tags characteristic of non-coding RNAs. , 2001 .

[22]  Wen-Hsiung Li,et al.  The K(A)/K(S) ratio test for assessing the protein-coding potential of genomic regions: an empirical and simulation study. , 2002, Genome research.

[23]  R. Lehmann,et al.  A Noncoding RNA Is Required for the Repression of RNApolII-Dependent Transcription in Primordial Germ Cells , 2004, Current Biology.