TSR1

gene
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Also known as FLJ10534

Summary

TSR1 (TSR1 ribosome maturation factor, HGNC:25542) is a protein-coding gene on chromosome 17p13.3, encoding Pre-rRNA-processing protein TSR1 homolog (Q2NL82). Required during maturation of the 40S ribosomal subunit in the nucleolus. It is a common-essential gene (DepMap: required in 100.0% of cancer cell lines).

Enables RNA binding activity. Predicted to be involved in endonucleolytic cleavage of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) and maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA). Predicted to be located in nucleolus.

Source: NCBI Gene 55720 — RefSeq curated summary.

At a glance

  • Gene–disease (curated): idiopathic spontaneous coronary artery dissection (Limited, GenCC)
  • GWAS associations: 6
  • Clinical variants (ClinVar): 140 total
  • Cancer dependency (DepMap): dependent in 100.0% of screened cell lines (common-essential)
  • MANE Select transcript: NM_018128

Identifiers

Gene identifiers

FieldValue
HGNC IDHGNC:25542
Approved symbolTSR1
NameTSR1 ribosome maturation factor
Location17p13.3
Locus typegene with protein product
StatusApproved
AliasesFLJ10534
Ensembl geneENSG00000167721
Ensembl biotypeprotein_coding
OMIM611214
Entrez55720

Gene structure

Transcript identifiers

Ensembl transcripts: 12 — 9 protein_coding, 2 nonsense_mediated_decay, 1 retained_intron

ENST00000301364, ENST00000571806, ENST00000575049, ENST00000576112, ENST00000576202, ENST00000909712, ENST00000915542, ENST00000915543, ENST00000915544, ENST00000915545, ENST00000947905, ENST00000947906

RefSeq mRNA: 1 — MANE Select: NM_018128 NM_018128

CCDS: CCDS32525

Canonical transcript exons

ENST00000301364 — 15 exons

ExonStartEnd
ENSE0000111610423245042324578
ENSE0000111610523305152330625
ENSE0000111610623344722334896
ENSE0000111610723293432329475
ENSE0000111610823246892324829
ENSE0000111611023321692332359
ENSE0000111611323309472331109
ENSE0000111611423335572333716
ENSE0000111611623329612333124
ENSE0000111611723360372336140
ENSE0000143434723223962324374
ENSE0000268303123363312336457
ENSE0000352544723253042325420
ENSE0000356224523352602335394
ENSE0000363959023355112335730

Expression profiles

Bgee: expression breadth ubiquitous, 300 present calls, max score 98.87.

FANTOM5 (CAGE): breadth ubiquitous, TPM avg 39.5427 / max 296.1890, expressed in 1816 samples.

FANTOM5 promoters (1 alternative TSS)

Promoter IDTPM avgSamples expressed
16382039.54271816

Top tissues by expression

300 total, by Bgee expression score (0-100, higher = more expressed):

TissueAnatomy IDExpression scoreQuality
cervix squamous epitheliumUBERON:000692298.87gold quality
nippleUBERON:000203098.21gold quality
mucosa of paranasal sinusUBERON:000503098.19gold quality
buccal mucosa cellCL:000233698.16gold quality
renal medullaUBERON:000036297.96gold quality
male germ cellCL:000001597.80gold quality
spermCL:000001997.79gold quality
cardia of stomachUBERON:000116297.78gold quality
ventral tegmental areaUBERON:000269197.78gold quality
superficial temporal arteryUBERON:000161497.71gold quality
lateral globus pallidusUBERON:000247697.65gold quality
olfactory bulbUBERON:000226497.59gold quality
superior surface of tongueUBERON:000737197.56gold quality
blood vessel layerUBERON:000479797.41gold quality
pylorusUBERON:000116697.35gold quality
pericardiumUBERON:000240797.27gold quality
subthalamic nucleusUBERON:000190697.17gold quality
inferior vagus X ganglionUBERON:000536397.10gold quality
lateral nuclear group of thalamusUBERON:000273697.05gold quality
periodontal ligamentUBERON:000826697.03gold quality
trigeminal ganglionUBERON:000167597.02gold quality
medulla oblongataUBERON:000189696.91gold quality
pharyngeal mucosaUBERON:000035596.89gold quality
saphenous veinUBERON:000731896.88gold quality
type B pancreatic cellCL:000016996.76gold quality
superior vestibular nucleusUBERON:000722796.73gold quality
tongueUBERON:000172396.70gold quality
substantia nigra pars compactaUBERON:000196596.68gold quality
body of tongueUBERON:001187696.66gold quality
vena cavaUBERON:000408796.54gold quality

Single-cell (SCXA)

Detected in 1 experiment(s), a significant marker in 1.

ExperimentMarker?Max mean expression
E-ANND-3no0.00

Regulation

Is transcription factor: no

miRNA regulators (miRDB)

107 targeting TSR1, top 30 by miRDB confidence (max_score; target_count = how many genes the miRNA targets in total — lower means more specific):

miRNAMax scoreAvg scoremiRNA target_count
HSA-MIR-4425100.0067.591049
HSA-MIR-10401-5P99.9965.79948
HSA-MIR-186-5P99.9970.833707
HSA-MIR-150-5P99.9966.691976
HSA-MIR-314899.9775.066478
HSA-MIR-3605-5P99.9667.12932
HSA-MIR-1250-3P99.9670.044038
HSA-MIR-6778-3P99.9667.292693
HSA-MIR-128-3P99.9571.172484
HSA-MIR-216A-3P99.9571.192505
HSA-MIR-130599.9171.433443
HSA-MIR-990299.8969.152250
HSA-MIR-3681-3P99.8870.462254
HSA-MIR-797899.8666.90856
HSA-MIR-313399.8170.923506
HSA-MIR-4766-5P99.7569.232662
HSA-MIR-453099.6966.471509
HSA-MIR-509399.6769.262291
HSA-MIR-6762-3P99.6666.941188
HSA-MIR-7157-5P99.6669.331829
HSA-MIR-3177-5P99.6570.381174
HSA-MIR-3679-3P99.6469.881599
HSA-MIR-4743-3P99.6268.122095
HSA-MIR-431099.5968.842527
HSA-MIR-1212299.5669.331672
HSA-MIR-6733-3P99.5467.801281
HSA-MIR-105-5P99.5469.242060
HSA-MIR-7853-5P99.5469.302055
HSA-MIR-103A-1-5P99.3967.781545
HSA-MIR-103A-2-5P99.3967.721577

Functional genomics

DepMap (CRISPR cell-line fitness): dependent in 100.0% of screened cell lines, common-essential.

Literature-anchored findings (GeneRIF, showing 3)

  • hTsr1 is involved downstream in nuclear export of the pre-40S particles. (PMID:20805244)
  • This study describes the clinical characteristics of the Chinese Han population with Spontaneous Coronary Artery Dissection and identified TSR1 as a potential causal gene, which might bring about a further progress in diagnosis and treatment of the disorder. (PMID:31296287)
  • a novel congenital cataract candidate gene TSR1 was identified by whole genome sequencing and Sanger sequencing in a Chinese congenital cataract family (PMID:32102773)

Cross-species orthologs

5 orthologs

OrganismSymbolGene ID
danio_reriotsr1ENSDARG00000007744
mus_musculusTsr1ENSMUSG00000038335
rattus_norvegicusTsr1ENSRNOG00000002980
drosophila_melanogasterTsr1FBGN0037073
caenorhabditis_elegansWBGENE00006497

Paralogs (1): BMS1 (ENSG00000165733)

Protein

Protein identifiers

Pre-rRNA-processing protein TSR1 homologQ2NL82 (reviewed: Q2NL82)

All UniProt accessions (4): Q2NL82, I3L1Q5, I3L4E8, I3L4T0

UniProt curated annotations — full annotation on UniProt →

Function. Required during maturation of the 40S ribosomal subunit in the nucleolus.

Subcellular location. Nucleus. Nucleolus.

Similarity. Belongs to the TRAFAC class translation factor GTPase superfamily. Bms1-like GTPase family. TSR1 subfamily.

RefSeq proteins (1): NP_060598* (*=MANE)

Domains & families (InterPro)

IDNameType
IPR007034BMS1_TSR1_CDomain
IPR012948AARP2CNDomain
IPR030387G_Bms1/Tsr1_domDomain
IPR039761Bms1/Tsr1Family

Pfam: PF04950, PF08142, PF22298

UniProt features (85 total): strand 48, helix 14, turn 12, sequence variant 4, sequence conflict 2, compositionally biased region 2, chain 1, domain 1, region of interest 1

Structure

Experimental structures (PDB)

17 structures.

PDBMethodResolution (Å)
6ZMTELECTRON MICROSCOPY3
7WTUELECTRON MICROSCOPY3
7WTZELECTRON MICROSCOPY3
7WTTELECTRON MICROSCOPY3.1
7WTXELECTRON MICROSCOPY3.1
6ZN5ELECTRON MICROSCOPY3.2
7WTSELECTRON MICROSCOPY3.2
7WTWELECTRON MICROSCOPY3.2
7WU0ELECTRON MICROSCOPY3.3
7WTVELECTRON MICROSCOPY3.5
6G18ELECTRON MICROSCOPY3.6
8ZDDELECTRON MICROSCOPY3.7
8ZDCELECTRON MICROSCOPY3.8
6G4SELECTRON MICROSCOPY4
6G51ELECTRON MICROSCOPY4.1
6G4WELECTRON MICROSCOPY4.5
6G53ELECTRON MICROSCOPY4.5

Predicted structure (AlphaFold)

ModelpLDDTFraction very-high
AF-Q2NL82-F180.620.48

Function

Pathways and Gene Ontology

Reactome pathways

1 pathways

IDPathway
R-HSA-6791226Major pathway of rRNA processing in the nucleolus and cytosol

MSigDB gene sets: 209 (showing top): GSE45365_NK_CELL_VS_CD11B_DC_DN, GSE45365_NK_CELL_VS_BCELL_UP, RNGTGGGC_UNKNOWN, GOBP_RIBOSOME_BIOGENESIS, SHEPARD_CRASH_AND_BURN_MUTANT_UP, GOBP_MATURATION_OF_SSU_RRNA, CACCAGC_MIR138, MONNIER_POSTRADIATION_TUMOR_ESCAPE_UP, PUJANA_CHEK2_PCC_NETWORK, AP1_Q4_01, MILI_PSEUDOPODIA_HAPTOTAXIS_UP, GOBP_RIBOSOMAL_SMALL_SUBUNIT_BIOGENESIS, GOBP_MATURATION_OF_SSU_RRNA_FROM_TRICISTRONIC_RRNA_TRANSCRIPT_SSU_RRNA_5_8S_RRNA_LSU_RRNA, GROSS_HYPOXIA_VIA_HIF1A_UP, TGANTCA_AP1_C

GO Biological Process (3): maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) (GO:0000462), endonucleolytic cleavage of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) (GO:0000479), ribosome biogenesis (GO:0042254)

GO Molecular Function (5): RNA binding (GO:0003723), GTPase activity (GO:0003924), GTP binding (GO:0005525), U3 snoRNA binding (GO:0034511), protein binding (GO:0005515)

GO Cellular Component (4): nucleoplasm (GO:0005654), nucleolus (GO:0005730), cytosol (GO:0005829), nucleus (GO:0005634)

Reactome top-level categories

Rollup of top-1 pathways:

CategoryPathways
rRNA processing in the nucleus and cytosol1

GO top-level categories

Rollup of top GO terms by namespace:

CategoryTerms
nuclear lumen2
cellular anatomical structure2
maturation of SSU-rRNA1
rRNA processing1
ribonucleoprotein complex biogenesis1
nucleic acid binding1
ribonucleoside triphosphate phosphatase activity1
guanyl ribonucleotide binding1
purine ribonucleoside triphosphate binding1
snoRNA binding1
binding1
intracellular membraneless organelle1
cytoplasm1
intracellular membrane-bounded organelle1

Protein interactions and networks

STRING

2538 interactions, top by confidence (×1000):

Protein AProtein BPartner UniProtScore
TSR1BYSLQ13895963
TSR1LTV1Q96GA3943
TSR1RIOK2Q9BVS4937
TSR1TSR2Q969E8902
TSR1RCL1Q9Y2P8892
TSR1PNO1Q9NRX1883
TSR1NOB1Q9ULX3877
TSR1TSR3Q9UJK0810
TSR1RIOK1Q9BRS2805
TSR1RRP12Q5JTH9744
TSR1POLR2BP30876674
TSR1MCM7P33993667
TSR1WDR12Q9GZL7665
TSR1RPS3P23396664
TSR1TXNL4BQ9NX01655

IntAct

146 interactions, top by confidence:

ABTypeScore
XPCCETN3psi-mi:“MI:0914”(association)0.730
PTK2TGFB1I1psi-mi:“MI:0914”(association)0.680
TSR1RPS3psi-mi:“MI:0914”(association)0.640
BYSLPARNpsi-mi:“MI:0914”(association)0.640
TSR1CIDEBpsi-mi:“MI:0915”(physical association)0.560
TSR1RHBDD2psi-mi:“MI:0915”(physical association)0.560
ZBTB48ZBTB24psi-mi:“MI:0914”(association)0.530
TSR1PARNpsi-mi:“MI:0914”(association)0.530
ABCE1EIF3Hpsi-mi:“MI:0914”(association)0.530
KPNB1POM121Cpsi-mi:“MI:0914”(association)0.530
RIOK2BYSLpsi-mi:“MI:0914”(association)0.530
TSR1RPSApsi-mi:“MI:0914”(association)0.530
LIN28BELAVL2psi-mi:“MI:0914”(association)0.530
H3C1SMCHD1psi-mi:“MI:2364”(proximity)0.410
TSR1H1-5psi-mi:“MI:0915”(physical association)0.400
Csnk1epsi-mi:“MI:0915”(physical association)0.400
SDC1ILVBLpsi-mi:“MI:0915”(physical association)0.400
TSR1YWHAZpsi-mi:“MI:0915”(physical association)0.400
PCNATSR1psi-mi:“MI:0915”(physical association)0.370
Rpl35RPS6psi-mi:“MI:0914”(association)0.350
Eif3aRPSApsi-mi:“MI:0914”(association)0.350
Cep78ING5psi-mi:“MI:0914”(association)0.350
RPL10RPS6psi-mi:“MI:0914”(association)0.350
Srp72psi-mi:“MI:0914”(association)0.350
CDK2AP1MTA3psi-mi:“MI:0914”(association)0.350
Rrbp1PIPSLpsi-mi:“MI:0914”(association)0.350
NOP56C12orf43psi-mi:“MI:0914”(association)0.350
IBTKPOP7psi-mi:“MI:0914”(association)0.350
HNRNPUpsi-mi:“MI:0914”(association)0.350

BioGRID (446): TSR1 (Affinity Capture-MS), TSR1 (Affinity Capture-MS), TSR1 (Affinity Capture-MS), TSR1 (Affinity Capture-MS), TSR1 (Affinity Capture-MS), HSP90B1 (Co-fractionation), LTV1 (Co-fractionation), MRPL2 (Co-fractionation), POLR1C (Co-fractionation), POLR1D (Co-fractionation), RPS7 (Co-fractionation), TSR1 (Co-fractionation), TSR1 (Co-fractionation), TSR1 (Co-fractionation), TSR1 (Co-fractionation)

ESM2 similar proteins: A3GFS1, A3LXF0, A5DBG1, A6ZMA9, A7TTC4, O13956, O74774, O74945, O94653, P09880, P14906, P23615, P25038, P32769, P33755, P36048, P41814, P53893, Q04660, Q07381, Q08965, Q09915, Q19329, Q2NL82, Q3MNT0, Q54JN0, Q54YA7, Q5A6Q4, Q5R434, Q5SWD9, Q5XGY1, Q61WR2, Q6BKK7, Q6BRG6, Q6BRJ9, Q6BVE1, Q6C7B9, Q6CF35, Q6CIH3, Q6CLF6

Diamond homologs: O13956, Q07381, Q2NL82, Q5R434, Q5SWD9, Q5XGY1, Q9VP47, Q19329, Q54YA7, Q61WR2

SIGNOR signaling

0 interactions.

Enriched among interaction partners

Reactome pathways and GO biological processes over-represented among this gene’s 183 IntAct physical interaction partners (hypergeometric vs the genome-wide background, BH-FDR, gene-set size 15–500, ranked by fold). A functional readout of the neighbourhood — distinct from this gene’s own memberships above, and biased toward well-studied / hub proteins, so read it as themes rather than proof.

Reactome pathways:

PathwayPartnersFoldFDR
Eukaryotic Translation Initiation922.1×9e-09
Cap-dependent Translation Initiation922.1×9e-09
SARS-CoV-1 modulates host translation machinery922.1×9e-09
Eukaryotic Translation Elongation919.9×2e-08
Activation of the mRNA upon binding of the cap-binding complex and eIFs, and subsequent binding to 43S919.4×2e-08
Formation of the ternary complex, and subsequently, the 43S complex1118.8×1e-09
Influenza Infection1318.1×6e-11
Influenza Viral RNA Transcription and Replication1017.1×1e-08

GO biological processes:

GO termPartnersFoldFDR
maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA)625.3×2e-05
NLS-bearing protein import into nucleus525.1×2e-04
ribosomal small subunit biogenesis1521.4×3e-13
ribosomal large subunit biogenesis616.6×2e-04
cytoplasmic translation1416.2×8e-11
negative regulation of innate immune response516.0×1e-03
rRNA processing1412.4×2e-09
translational initiation511.2×5e-03

Disease & clinical

Clinical variants and AI predictions

ClinVar

140 variants total. Per-class counts are floors (≥ shown; pagination cap):

ClassificationCount (floor)
Pathogenic0
Likely pathogenic0
Uncertain significance111
Likely benign6
Benign4

Top pathogenic / likely-pathogenic (0)

SpliceAI

1887 predictions. Top by Δscore:

VariantEffectΔscore
17:2323133:CAG:Cacceptor_loss1.0000
17:2323134:A:Cacceptor_loss1.0000
17:2324371:GTAC:Gacceptor_gain1.0000
17:2324372:TAC:Tacceptor_gain1.0000
17:2324372:TACC:Tacceptor_loss1.0000
17:2324374:CCTAG:Cacceptor_loss1.0000
17:2324375:C:CAacceptor_loss1.0000
17:2324375:C:CCacceptor_gain1.0000
17:2324576:CCTCT:Cacceptor_gain1.0000
17:2324580:T:Cacceptor_gain1.0000
17:2324580:T:TCacceptor_gain1.0000
17:2324587:C:CTacceptor_gain1.0000
17:2324594:A:Cacceptor_gain1.0000
17:2324712:A:ACdonor_gain1.0000
17:2324713:C:CCdonor_gain1.0000
17:2324713:CTA:Cdonor_gain1.0000
17:2324825:CATTC:Cacceptor_gain1.0000
17:2324827:TTC:Tacceptor_gain1.0000
17:2324828:TCCTA:Tacceptor_loss1.0000
17:2324830:C:CCacceptor_gain1.0000
17:2324830:C:CGacceptor_loss1.0000
17:2324831:T:Gacceptor_loss1.0000
17:2325297:AACTT:Adonor_loss1.0000
17:2325298:ACTTA:Adonor_loss1.0000
17:2325301:TA:Tdonor_loss1.0000
17:2325302:A:ATdonor_loss1.0000
17:2325425:CA:Cacceptor_gain1.0000
17:2325426:A:ACacceptor_gain1.0000
17:2325432:G:Cacceptor_gain1.0000
17:2325432:G:GCacceptor_gain1.0000

AlphaMissense

5327 scored. Top likely-pathogenic:

VariantProtein changeam_pathogenicity
17:2324365:C:TG749D0.999
17:2324524:C:TG739E0.999
17:2324748:C:TG701D0.999
17:2331066:A:GW514R0.999
17:2331066:A:TW514R0.999
17:2324324:C:GD763H0.998
17:2324352:G:CC753W0.998
17:2324525:C:GG739R0.998
17:2324525:C:TG739R0.998
17:2324548:A:GL731P0.998
17:2324709:C:GR714P0.998
17:2324323:T:AD763V0.997
17:2324323:T:GD763A0.997
17:2324354:A:GC753R0.997
17:2324365:C:AG749V0.997
17:2324562:A:CF726L0.997
17:2324562:A:TF726L0.997
17:2324564:A:GF726L0.997
17:2324576:C:GD722H0.997
17:2324700:A:GF717S0.997
17:2324749:C:GG701R0.997
17:2324750:A:CS700R0.997
17:2324750:A:TS700R0.997
17:2324752:T:GS700R0.997
17:2332178:G:TA496D0.997
17:2332213:A:CF484L0.997
17:2332213:A:TF484L0.997
17:2332215:A:GF484L0.997
17:2324305:A:GL769P0.996
17:2324317:A:TV765E0.996

dbSNP variants (sampled 300 via entrez): RS1000210601 (17:2322271 G>GT), RS1000493871 (17:2325983 A>G), RS1000566864 (17:2325696 T>A), RS1000579269 (17:2322655 C>T), RS1000724567 (17:2331763 T>C), RS1000822271 (17:2328000 C>T), RS1000840496 (17:2335114 T>C), RS1000856020 (17:2337391 C>G), RS1001062259 (17:2328342 T>C), RS1001158695 (17:2326830 C>T), RS1001377510 (17:2326977 C>T), RS1001388705 (17:2329057 C>T), RS1001529402 (17:2334739 G>A), RS1001723141 (17:2333596 G>A), RS1002005500 (17:2338237 G>A,C)

Disease associations

OMIM: gene MIM:611214 | disease phenotypes:

GenCC curated gene-disease

DiseaseClassificationInheritance
idiopathic spontaneous coronary artery dissectionLimitedAutosomal dominant

Mondo (1): idiopathic spontaneous coronary artery dissection (MONDO:0007385)

Orphanet (0):

HPO phenotypes

0 total (0 of 0 shown, HPO-id order):

GWAS associations

6 associations (top):

StudyTraitp-value
GCST000442_1Aortic root size2.000000e-11
GCST002539_85Schizophrenia3.000000e-10
GCST004521_272Autism spectrum disorder or schizophrenia7.000000e-10
GCST006803_64Schizophrenia5.000000e-10
GCST010703_278Brain morphology (MOSTest)2.000000e-15
GCST90000025_116Appendicular lean mass4.000000e-16

EFO canonical traits (2, from GWAS)

EFO IDTrait name
EFO:0004346neuroimaging measurement
EFO:0004980appendicular lean mass

MeSH disease descriptors (1)

DescriptorNameTree numbers
C565153Coronary Artery Dissection, Spontaneous (supp.)

Drugs & pharmacology

Drug and pharmacology data

Is drug target: no

PharmGKB: 1 entry (VIP=true, CPIC=false)

CTD chemical–gene interactions

42 total (human), top 30 by PubMed support.

ChemicalActions (top 5)PubMed papers
bisphenol Adecreases expression3
Tretinoindecreases expression3
sodium arsenitedecreases expression2
potassium chromate(VI)affects cotreatment, increases expression2
Nickelincreases expression2
Valproic Acidincreases expression, affects expression2
Cyclosporineincreases expression2
Aflatoxin B1increases expression, increases methylation2
TAK-243increases sumoylation1
alpha phellandrenedecreases expression1
triphenyl phosphateaffects expression1
sodium arsenatedecreases expression1
perfluorooctanoic acidincreases expression1
epigallocatechin gallateaffects cotreatment, increases expression1
chromium hexavalent ionincreases expression1
4-phenylbutyric aciddecreases expression1
cylindrospermopsinincreases expression1
4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamideaffects cotreatment, decreases expression1
LDN 193189affects cotreatment, decreases expression1
Resveratrolaffects cotreatment, increases expression1
Sunitinibincreases expression1
Arsenicaffects methylation1
Benzo(a)pyreneincreases methylation1
Dichlorodiphenyl Dichloroethyleneincreases expression1
Dexamethasonedecreases expression1
Dimethyl Sulfoxideincreases expression1
Doxorubicinincreases expression1
Enzyme Inhibitorsdecreases activity, increases O-linked glycosylation1
Estradiolincreases expression1
Ethyl Methanesulfonatedecreases expression1

Clinical trials (associated diseases)

0 trials via MONDO — disease-level, not drug-specific.