(RNA-) Structuredness in Viruses

Roman Ochsenreiter

TBI WienUniversity of Vienna

16.2.2017 / TBI Winterseminar

Roman Ochsenreiter (RNA-)Structuredness in Viruses

Viruses IMinimal Life Forms

Obligate intracellularparasites

No organelles

(Very) small size

DNA/RNA genomes

Electron micrograph of an Ebola Virion, size: 1000 nm(image source: wikipedia)

Roman Ochsenreiter (RNA-)Structuredness in Viruses

Viruses IIMinimal Life Forms

Figure: Crystal Structure and Schematic of Rhinovirus A 1

Genome size (nt): Human: 3.2 × 109 Rhinovirus: 7 × 103

1Zhao et al. 1996Roman Ochsenreiter (RNA-)Structuredness in Viruses

Viruses and RNA Structure

Viruses have tightly packed genomesSpecific Functions are often provided by RNA Structures

Figure: Pestivirus IRES1, Rhinovirus CRE-Element

1Rfam IDs: RF00209, RF00220Roman Ochsenreiter (RNA-)Structuredness in Viruses

Viral DiversityBaltimore Classification

Roman Ochsenreiter (RNA-)Structuredness in Viruses

Viral DiversityBaltimore Classification

Roman Ochsenreiter (RNA-)Structuredness in Viruses

Viral DiversityViral Phylogeny

1957. 79 Kokober a_vi r us3165. 21 Banzi _vi r us_st r ai n_SAH_336_pol ypr ot ei n_gene3106. 79 Edge_Hi l l _vi r us_st r ai n_YMP_48_pol ypr ot ei n_gene

645. 80

913. 37 Wessel sbr on_vi r us952. 63 Sepi k_vi r us

2326. 73

3252. 77 Yel l ow_f ever _vi r us

303. 82

753. 11

1588. 25 Sokol uk_vi r us_st r ai n_LEI V- 400K1624. 75 Ent ebbe_bat _vi r us

1044. 84

2686. 66 Yokose_vi r us

2225. 58

226. 98

4589. 77 Par ai so_Escondi do_vi r us_i sol at e_Ecuador 2012

431. 84

2329. 70 Par r amat t a_Ri ver _vi r us_i sol at e_92- B1157452296. 30 Hanko_vi r us_pol ypr ot ei n_gene

3246. 44

2703. 05 Aedes_f l avi vi r us_genomi c_RNA2650. 95 Kami t i _Ri ver _vi r us

2955. 81

619. 84

6323. 23 Mer cadeo_vi r us_i sol at e_ER- M10599. 73 Quang_Bi nh_vi r us593. 27 Mosqui t o_f l avi vi r us_i sol at e_LSFl avi V- A20- 09

2128. 45

2700. 05 Cul ex_f l avi vi r us

3303. 02

501. 91

2744. 36

11426. 33 Tamana_bat _vi r us

2278. 26

325. 03Spani sh_goat _encephal i t i s_vi r us_i sol at e_Ast Goat I REC2011445. 97 Loupi ng_i l l _vi r us

90. 87

411. 63 Ti ck- bor ne_encephal i t i s_vi r us

381. 34

795. 91 Omsk_hemor r hagi c_f ever _vi r us

448. 57

1274. 93 Langat _vi r us

435. 04

1681. 96 Al khur ma_vi r us

263. 67

1951. 24 Powassan_vi r us

489. 77

2494. 68 Kar shi _vi r us

1599. 66

2148. 99 Kama_vi r us_st r ai n_LEI V- 20776Tat _pol ypr ot ei n_gene2085. 01 Tyul eni y_vi r us_st r ai n_LEI V- 6C_pol ypr ot ei n_gene

1812. 66

1237. 18

2929. 92 J ut i apa_vi r us_st r ai n_J G- 1282982. 08 Modoc_vi r us

859. 26

3670. 74 Ri o_Br avo_vi r us

433. 36

4304. 39 Apoi _vi r us

1962. 39

472. 20

709. 24

652. 03

2786. 86 Chaoyang_vi r us2778. 14 Donggang_vi r us

1763. 85

633. 00

451. 44 West _Ni l e_vi r us_l i neage_1465. 56 West _Ni l e_vi r us_l i neage_2

1381. 70

1314. 02 Mur r ay_Val l ey_encephal i t i s_vi r us1354. 46 J apanese_encephal i t i s_vi r us1311. 54 Usut u_vi r us34. 98

499. 30

422. 01

2331. 87 Caci pacor e_vi r us_st r ai n_BeAn_3276000

529. 11

2575. 20 St . _Loui s_encephal i t i s_vi r us

174. 44

854. 63 Bagaza_vi r us783. 37 Nt aya_vi r us_i sol at e_I PDI A

603. 37

1423. 13 Tembusu_vi r us_st r ai n_J S804

1563. 28

3020. 28 I l heus_vi r us

1004. 46

41. 57

2519. 16 Dengue_vi r us_42253. 04 Dengue_vi r us_2

1686. 38 Dengue_vi r us_11693. 62 Dengue_vi r us_3

594. 96328. 09

2010. 88

150. 08

2001. 12 Spondweni _vi r us2055. 88 Zi ka_vi r us

1381. 72

3926. 61 Kedougou_vi r us

276. 39

3689. 41 Bussuquar a_vi r us

1957. 79 Spondveni (Zika) Virus

Dengue Virus

West Nile Virus / Japan Encephalitis Virus

Tick-borne Flaviviruses

Mosquito-borne FVs

Yellow Fever Virus

Figure: Picorna-like Viruses (left)1, Flavivirus (right

1Fauquet, Schrock; 2006Roman Ochsenreiter (RNA-)Structuredness in Viruses

Viral DiversityViral Phylogeny

Phylogenetic Tree of Viruses

GrüneFilamentöse

BakterienSpirochäten

Gram-positive

Proteobakterien

Cyanobakterien

Planctomyceten

BacteroidesCytophaga

Thermotogae

Aquifex

HalophileMethanosarcina

Methanobacterium

Methanococcus

T. celer

ThermoproteusPyrodicticum

Entamoebae Schleim-pilze

TierePilze

Pflanzen

Wimperntierchen

Flagellaten

Trichomonaden

Microsporidien

Diplomonaden

Roman Ochsenreiter (RNA-)Structuredness in Viruses

Virus Ortholog Groups

Viral Clusters of Ortholog Groups

Groups of Ortholog proteinsCalculated on Protein alignmentsBased on newest RefSeq Virus GenomesIncludes all annotated viral species

Roman Ochsenreiter (RNA-)Structuredness in Viruses

Virus Ortholog Groups

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40 60 80 100

0.0

0.2

0.4

0.6

0.8

1.0

Viral Ortholog Proteins (mRNA) MPI vs SCI

MPI

SC

I

Roman Ochsenreiter (RNA-)Structuredness in Viruses

Virus Ortholog Groups

Viral Ortholog Proteins (mRNA) SCI

SCI

Cou

nts

0.0 0.2 0.4 0.6 0.8 1.0

020

0040

0060

0080

0010

000

1200

0

Viral Ortholog Proteins (mRNA) MPI

MPI

Cou

nts

20 40 60 80 1000

1000

2000

3000

4000

5000

Roman Ochsenreiter (RNA-)Structuredness in Viruses

Viral Ortholog ProteinsConserved sub-alignments

10 20 30 40

- - - - - - - - A TGGG T A A TG T TGC A AC TC T T A T A A AC A AGGC T A T T- - - - - - - - A TGGG T AG T T A TGC T AG T A T T T TC T A TC A TGC T T TG- - - - - - - - A TGGG T AG T T A TGC T AG T A T T T T T T A TC AC GC AC T A- - - - - - - - A TGGG A TC AG TG TGG AG T A T AG TG AG TC AGGC TG T A- - - - - - - - A TGGG T AG TG TG TGG TC TC AC A TC TC G AC C GC C T TC- - - - - - - - A TGGG T AGC A T A TC A TC C C AC A T TG TC AC TGC C T TC- - - - - - - - A TGGG T AGC C T T TGG TC C A AG A T TGGC C AGC TC T T TTGGGG TC C A TGC A A AGC C T T T T TG AC A A A A T TGGC C A AC T T T T T- - - - - - - - A TGGG T TC T A T AC TC AC C C AC A TC AC T AC AGC C T TC

10 20 30 40

- - - - - - - - - A TGGG T A A TG T TGC A AC TC T T A T A A AC A AGGC T A T T- - - - - - - - - A TGGG T AG T T A TGC T AG T A T T T TC T A TC A TGC T T TG- - - - - - - - - A TGGG T AG T T A TGC T AG T A T T T T T T A TC AC GC AC T A- - - - - - - - - A TGGG A TC AG TG TGG AG T A T AG TG AG TC AGGC TG T A- - - - - - - - - A TGGG T AG TG TG TGG TC TC AC A TC TC G AC C GC C T TC- - - - - - - - - A TGGG T AGC A T A TC A TC C C AC A T TG TC AC TGC C T TC

- - - - - - - - - A TGGG T AGC C T T TGG TC C A AG A T TGGC C AGC TC T T TA TGGGG TC C A TGC A A AGC C T T T T TG AC A A A A T TGGC C A AC T T T T T- - - - - - - - - A TGGG T TC T A T AC TC AC C C AC A TC AC T AC AGC C T TC

SCI = 0.0

SCI = 0.9

SCI = 0.1

No overall consensus structure but. . .Strong structure conservation in subparts of the alignment!

Roman Ochsenreiter (RNA-)Structuredness in Viruses

Viruses and structured RNAsMore than UTRs. . .

The ProblemViruses are a highly diverse group of organismsJust a fraction is investigated for ncRNAsUTRs get the most attention

AimsScreen all viruses for structured RNAsTake a close look at CDS regions

MethodsRNAzR-scape

Roman Ochsenreiter (RNA-)Structuredness in Viruses

DatasetIsolation Pipeline

1 Fetch Species from NCBItaxonomy database

2 Fetch Refseq Sequences

3 Fetch genomes from NCBINucleotide Database

4 Align with clustal Omega

5 Multiple consistencychecks in-between

Name #SpeciesNCBI Tax DB 18939Refseq DB 8299Consistent Species 1580

Type #SpeciesdsDNA 422ssDNA 276dsRNA 142ssRNA 600

RT 81Satellites 32

Roman Ochsenreiter (RNA-)Structuredness in Viruses

Results

Significant Hit size

Locus Length

Cou

nts

0 200 400 600 800 1000 1200

050

0010

000

1500

020

000

2500

0

Roman Ochsenreiter (RNA-)Structuredness in Viruses

Results

0 50000 100000 150000 200000

010

020

030

040

050

0

Virus Genome Length

#No

of R

NAz

Hits

#RNAz hits vs. Genome Length

Roman Ochsenreiter (RNA-)Structuredness in Viruses

Results

#Hits w/ Cov (G) w/ Cov (RAF)Total Hits 73564 4216 43554

CDS 69941 4806 41579UTR 3664 410 1975

Roman Ochsenreiter (RNA-)Structuredness in Viruses

Summary & Outlook

SummaryStructured Elements are widespread in VirusesLots of Covariation in most elements!CDS and UTRs are equally affected

OutlookCheck again with structure-based alignmentsBuild covariance models for all significant lociSearch for ’structural orthologs’

Roman Ochsenreiter (RNA-)Structuredness in Viruses

Acknowledgements

Thanks to. . .

Ivo HofackerMichael Kiening (LMU Munich)Dimitrji Frishman (LMU Munich)Hans-Jörg Hellinger (Uni Wien)Thomas Rattei (Uni Wien)

. . . and you!

Roman Ochsenreiter (RNA-)Structuredness in Viruses