(RNA-) Structuredness in Viruses...Viruses I Minimal Life Forms Obligate intracellular parasites No...

(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)


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


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


Virus Ortholog Groups

Viral Clusters of Ortholog Groups

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



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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



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


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!


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


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


Results

Significant Hit size

Locus Length

Cou

nts

0 200 400 600 800 1000 1200

050

0010

000

1500

020

000

2500

0


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


Results

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

CDS 69941 4806 41579UTR 3664 410 1975


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’


Acknowledgements

Thanks to. . .

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

. . . and you!


(RNA-) Structuredness in Viruses...Viruses I Minimal Life Forms Obligate intracellular parasites No...

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