Systematic identification of non-coding RNA 2,2,7-trimethylguanosine cap structures in Caenorhabditis elegans.
BACKGROUND
The 2,2,7-trimethylguanosine (TMG) cap structure is an important functional characteristic of ncRNAs with critical cellular roles, such as some snRNAs. Here we used immunoprecipitation with both K121 and R1131 anti-TMG antibodies to systematically identify the TMG cap structures for all presently characterized ncRNAs in C. elegans.


RESULTS
The two anti-TMG antibodies precipitated a similar group of the C. elegans ncRNAs. All snRNAs known to have a TMG cap structure were found in the precipitate, indicating that our identification system was efficient. Other ncRNA families related to splicing, such as SL RNAs and Sm Y RNAs, were also found in the precipitate, as were 7 C/D box snoRNAs. Further analysis showed that the SL RNAs and the Sm Y RNAs shared a very similar Sm binding site element (AAU4-5GGA), which sequence composition differed somewhat from those of other U snRNAs. There were also 16 ncRNAs without an Sm binding site element in the precipitate, suggesting that for these ncRNAs, TMG formation may occur independently of Sm proteins.


CONCLUSIONS
Our results showed that most ncRNAs predicted to be transcribed by RNA polymerase II had a TMG cap, while those predicted to be transcribed by RNA plymerase III or located in introns did not have a TMG cap structure. Compared to ncRNAs without a TMG cap, TMG-capped ncRNAs tended to have higher expression levels. Five functionally non-annotated ncRNAs also have a TMG cap structure, which might be helpful for identifying the cellular roles of these ncRNAs.
Jia
Dong
D
Bioinformatics Laboratory, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China. donging@gmail.com
Cai
Lun
L
He
Housheng
H
Skogerbø
Geir
G
Li
Tiantian
T
Aftab
Muhammad Nauman
MN
Chen
Runsheng
R
D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000494:pharmacology; D001483:Base Sequence; D001665:Binding Sites; D017173:Caenorhabditis elegans / Q000235:genetics / Q000378:metabolism*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects; D047468:Immunoprecipitation; D008969:Molecular Sequence Data; D012315:RNA Caps / Q000096:biosynthesis / Q000737:chemistry* / Q000276:immunology; D022661:RNA, Untranslated / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D038621:Regulatory Sequences, Ribonucleic Acid; D014158:Transcription, Genetic / Q000187:drug effects
D000906:Antibodies; D012315:RNA Caps; D022661:RNA, Untranslated; D038621:Regulatory Sequences, Ribonucleic Acid