lncrnas long, non-coding rnas - units.it€¦ · long,non-coding rnas. mainly transcribed by rna...
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lncRNAs
long,non-codingRNAs
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MainlytranscribedbyRNAPol II
CharacteristicsoflncRNAs
Nuclearand/orcytoplasmaticlocaliztion
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DBE-T
lncRNAsinphysiology
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COMPARINGmRNAs- lncRNAs
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COMPARINGmRNAs- lncRNAs
lncRNAscanbe1. Intergenic (lincRNA):Donotintersectwithproteincodinggene
2.Genic:Intersectaproteincodinggene- Exonic- Intronic- overlapping
Divergent
Convergent
samestrandP P
P
PP
P
Closest protein coding gene lncRNAP:promoter Why? Why? Why?
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COMPARINGmRNAs- lncRNAs
A.LncRNAsdonothavecodingpotential:à LongestpossibleORFwassearchedinmRNA/lncRNAs
B.CAGEtagscanbefoundinlncRNAsà definedtranscriptionalstartsiteCAGEtagfreqeuncy increaseswithincreasedlncRNAexpressionlevels.mRNAsarecharacterizedbymoreCAGEtagsà LncRNAsaresimilartomRNAs
CAGE
freqeuncy
Expressionlevels
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COMPARINGmRNAs- lncRNAs
SPLICING:- 98%oflncRNAsarespliced,- RelevantSplice-siteprerequisitesatsplicedonor/acceptorareconserved- RemarkabletendencyoflncRNAs tohaveonly2exons(42%,mRNAs:6%)
BUT:LncRNAscontainfewerINTRONs!!mostlncRNAhaveonly1intron!!!;mRNAs3-6
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Introns/Exons fromlncRNAsareslightlylonger
COMPARINGmRNAs- lncRNAs
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COMPARINGmRNAs- lncRNAs
- lncRNAs areonaveragemuchshorter:ca.500nt- mRNAsarelongerandhavewidersizedistribution
- lncRNAs areuniformà littlealternativesplicing- mRNAs:largevarietyofalternativesplicing
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COMPARINGmRNAs- lncRNAs
conservation conservation conservation
EXONS:mRNA:highconservationlncRNA:reducedconservationBut: conservationishigherthanmRNAintron conservation
INTRONS:mRNA:higherconservationthanlncRNAs
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EvolutionalconservationoflncRNAs
BLASTofhumanlncRNAs against all availablemammalian genomes andidentify homologousgenes inother species:30%oflncRNA transcripts (n =4546)appear tobeprimatespecific.
Atotal of0.7% (101)oftranscriptsappear tobespecific tothehumanlineage.Asimilar number (134;1.0%)is foundinall ofthe18species analyzed here.
lncRNA
transcrip
ts
species
MostlncRNAtranscriptsareconservedin4-6species
lncRNAs showrelevantspeciesspecificity
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COMPARINGmRNAs- lncRNAsChromatinsignaturesattranscriptionalstartsitesareconservedbetweenmRNAsAndlncRNAs:MOSTlncRNAsARETRANSCRIBEDBYRNAPol II
Notshown:RNAPolIIpeaksattranscriptionstartsiteofmostlncRNAs
mRNAlncRNA 5psupportlncRNA
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COMPARINGmRNAs- lncRNAs
AbsoluteexpressionoflncRNAsismuchlowerthanmRNAexpression
RNA-seq
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COMPARINGmRNAs- lncRNAs
Microarry
AbsoluteexpressionoflncRNAsismuchlowerthanmRNAexpression
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COMPARINGmRNAs- lncRNAs
mRNAsshowbroadexpressionindifferenttissues–65%ofmRNAsareexpressedin16tissues
lncRNAs showtissue/cellsoecifcexpression–50%oflncRNAsexpressedin4-5tissues.
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lncRNAs locatemorefrequentlytochromatinthanmRNAs
lncRNAs locatemorefrequentlytothenucleusthanmRNAs
DeterminationoftranscriptabundanceNucleus/cytoplasma
DeterminationoftranscriptabundanceChromatin/nucleus
Data:RNAseq ofRNAslocatedinthenucleoplasmaandstickingtochromatin
Data:RNAseqofRNAslocatedinthenucleoplasmaandcytoplasma
COMPARINGmRNAs- lncRNAs
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COMPARINGmRNAs- lncRNAs
lncRNAexonsHavearemarkablePotentialtoencodefunctionalsmallRNAs(sRNAs)HIGHERTHANEXONSOFmRNAS!!!!snoRNA:Small nucleolar RNAs (snoRNAs) areaclassofsmall RNAmolecules thatprimarilyguide chemical modifications ofother RNAs,mainlyribosomal RNAs, transfer RNAs andsmallnuclear RNAs.snRNA: Smallnuclear ribonucleic acid(snRNA), alsocommonly referred toasU-RNA, isaclassofsmall RNAmolecules thatarefound within the splicing specklesandCajal bodies ofthecell nucleus ineukaryoticcells.tRNA: translationmiscRNAs: MiscRNA isshort formiscellaneous RNA, ageneral term foraseries ofmiscellaneous smallRNA.Itservesavarietyoffunctions, including some enzyme-likecatalysis andprocessing RNA after it isformed. Besides, some ofthesesmall RNAsmayserveasswitches.
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COMPARINGmRNAs- lncRNAs
lncRNAsrepresentabigclassoffunctionalelementsthat- controlledgeneexpression- areprocessed- lackproteincodingpotential- definedlocalization- frequentlyencodesRNAs- lowoverallconservation- 35%oflncRNAsareprimatespecific- expressionisratherlow– butcontrolled!- mostlytranscribedbyRNAPolII
FUNCTION:FORTHEVASTMAJORITYOFlncRNAsTHEBIOLOGICALFUNCTIONISUNKNOWN!!!!