Affinage

FAM98B

tRNA-splicing ligase complex subunit FAM98B · UniProt Q52LJ0

Length
433 aa
Mass
45.5 kDa
Annotated
2026-06-09
14 papers in source corpus 11 papers cited in narrative 11 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 4/4 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

FAM98B is a core structural subunit of the human tRNA ligase complex (tRNA-LC), an RTCB (HSPC117)-based assembly required for tRNA splicing and RNA ligation (PMID:24870230). Within this complex, FAM98B and CGI-99 associate through their N-terminal domains to form a stable co-folded heterodimer, and their C-terminal alpha-helical regions, together with DDX1, build the helical bundle that contacts the catalytic subunit RTCB and is required for complex integrity [PMID:34854379, PMID:bio_10.1101_2025.08.01.668197]. The FAM98B-CGI-99 heterodimer clamps the adaptor Ashwin in a pincer-like arrangement, and Ashwin engagement is specific to the FAM98B-containing complex, directing it to the nucleus for pre-tRNA splicing; the paralogous FAM98A- and FAM98C-containing complexes lack Ashwin and remain cytoplasmic, defining FAM98B as the determinant of the nuclear tRNA-biogenesis form [PMID:bio_10.1101_2025.08.01.668197, PMID:bio_10.1101_2025.08.01.668163]. FAM98B carries an exceptionally glycine-rich intrinsically disordered region that, in GGC repeat expansion disorders, is sequestered and depleted by polyglycine aggregates, disrupting tRNA-LC function and tRNA processing; Fam98b depletion in mice produces progressive motor coordination deficits and hindbrain pathology (PMID:40674500). Beyond the tRNA-LC, FAM98B co-purifies with DDX1, RTCB, and hCLE (C14orf166) in a cap-binding complex that shuttles between nucleus and cytoplasm and modulates mRNA translation (PMID:24608264, PMID:30833903), supports PRMT1 expression and proliferation in colorectal cancer cells redundantly with FAM98A (PMID:28040436), acts as a transcriptional activator at a TGFB1 regulatory variant (PMID:32757134), and is required for lysosome trafficking and bone resorption in osteoclasts (PMID:39857276).

Mechanistic history

Synthesis pass · year-by-year structured walk · 10 steps
  1. 2014 High

    Established FAM98B as a constituent of the RTCB-based tRNA splicing ligase complex, placing it in the machinery for tRNA splicing and RNA ligation.

    Evidence Biochemical purification and co-purification with RTCB

    PMID:24870230

    Open questions at the time
    • Did not define FAM98B's specific role within the complex
    • No structural arrangement of subunits resolved
  2. 2014 Medium

    Showed FAM98B participates in a transcription-dependent, nucleocytoplasmic-shuttling complex with DDX1, RTCB, and hCLE, extending its associations beyond tRNA ligation.

    Evidence Nuclear/cytoplasmic fractionation, reciprocal co-IP, photoactivatable GFP live imaging, and RNA silencing

    PMID:24608264

    Open questions at the time
    • Relationship between this shuttling complex and the tRNA-LC not resolved
    • Mechanism coupling FAM98B nuclear import to active transcription unknown
  3. 2016 Medium

    Linked the FAM98B/DDX1/C14orf166 complex to PRMT1 expression and cancer cell proliferation, and showed FAM98A and FAM98B act in the same pathway axis.

    Evidence Co-IP, shRNA knockdown, proliferation/colony assays, and double-knockdown epistasis in colorectal cancer cells

    PMID:28040436

    Open questions at the time
    • Molecular mechanism connecting the complex to PRMT1 expression unknown
    • Direct vs indirect regulation not distinguished
  4. 2019 Medium

    Demonstrated the FAM98B-containing complex binds the mRNA cap and modulates translation, assigning a post-transcriptional gene-expression role.

    Evidence Cap-analog affinity pulldown, competition/elution, RNA silencing, and polysome analysis

    PMID:30833903

    Open questions at the time
    • Whether FAM98B contacts the cap directly or via partners unresolved
    • Transcript selectivity of translational modulation not defined
  5. 2020 Medium

    Identified FAM98B as a transcriptional activator acting at a TGFB1 regulatory variant, a chromatin-associated role distinct from RNA ligation.

    Evidence DNA pull-down with mass spectrometry, luciferase reporter assay, and siRNA knockdown

    PMID:32757134

    Open questions at the time
    • Not independently replicated
    • Mechanism of sequence-specific DNA engagement and co-activator recruitment unknown
  6. 2021 High

    Resolved the architecture of the human tRNA-LC core, showing FAM98B and CGI-99 form an N-terminal heterodimer and contribute C-terminal helices required for complex integrity around RTCB.

    Evidence Crystal structure of RTCB and the CGI-99 N-terminal domain combined with deletion mutagenesis of inter-subunit interactions

    PMID:34854379

    Open questions at the time
    • Full-length FAM98B structure not determined
    • How the assembly is targeted to substrates not addressed
  7. 2025 High

    Defined the disease relevance of FAM98B's glycine-rich IDR, showing polyglycine aggregates sequester FAM98B to disrupt tRNA processing and cause neurological phenotypes in vivo.

    Evidence Biochemical aggregation assays, patient tissue analysis, FISH for tRNA splicing intermediates, and in vivo mouse knockdown with behavioral/histological readouts

    PMID:40674500

    Open questions at the time
    • Whether IDR sequestration is the sole disease driver in GGC disorders not established
    • Cell-type basis of hindbrain vulnerability not defined
  8. 2025 High

    Showed FAM98B determines the nuclear, Ashwin-bound form of the tRNA-LC dedicated to pre-tRNA splicing, distinguishing it functionally from FAM98A/C-containing cytoplasmic complexes.

    Evidence Cryo-EM structure, structure-based and NLS-disruption mutagenesis, fractionation, tRNA splicing intermediate assays, and RTCB-NLS rescue (preprints)

    PMID:bio_10.1101_2025.08.01.668163 PMID:bio_10.1101_2025.08.01.668197

    Open questions at the time
    • Determinants of paralog selection between FAM98A/B/C not defined
    • Peer-reviewed confirmation of the preprint findings pending
  9. 2025 Medium

    Extended FAM98B's cellular roles to osteoclast biology, showing it is required for lysosome trafficking and bone resorption with FAM98A redundancy.

    Evidence shRNA knockdown in osteoclast precursors, lysosomal trafficking and bone resorption assays, and qPCR for compensatory expression

    PMID:39857276

    Open questions at the time
    • Mechanism linking FAM98B to lysosome trafficking unknown
    • Whether this role depends on tRNA-LC activity not tested
  10. 2025 Medium

    Cross-species structure of the five-subunit tRNA-LC revealed a mobile DDX1 helicase module that can modulate RTCB activity, framing the regulatory dynamics of the FAM98B-containing assembly.

    Evidence Cryo-EM of Danio rerio tRNA-LC and thiol-based chemical crosslinking

    PMID:40220997

    Open questions at the time
    • Specific contribution of FAM98B to catalytic regulation not individually dissected
    • Functional consequence of DDX1 mobility on substrate handling not quantified

Open questions

Synthesis pass · forward-looking unresolved questions
  • How FAM98B's distinct activities — nuclear tRNA splicing, cap-binding translation, DNA-associated transcriptional activation, and osteoclast lysosome trafficking — are mechanistically coordinated within or independent of the RTCB complex remains unresolved.
  • No mechanism unifying the chromatin/transcriptional role with the RNA-processing complex
  • Whether non-tRNA-LC functions require complex assembly is untested

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0005198 structural molecule activity 3 GO:0003723 RNA binding 1 GO:0140110 transcription regulator activity 1
Localization
GO:0005634 nucleus 2 GO:0005829 cytosol 2
Pathway
R-HSA-8953854 Metabolism of RNA 3 R-HSA-74160 Gene expression (Transcription) 1
Complex memberships
cap-binding mRNA transport/translation complextRNA ligase complex (tRNA-LC)

Evidence

Reading pass · 11 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2014 FAM98B is a component of the human tRNA splicing ligase complex (tRNA-LC), which contains RTCB (HSPC117) as the catalytic subunit. FAM98B was identified as part of this multimeric complex required for tRNA splicing and RNA ligation. Biochemical purification and identification of complex components; co-purification with RTCB Nature High 24870230
2014 FAM98B co-purifies with DDX1, HSPC117 (RTCB), and hCLE (C14orf166) in both nuclear and cytoplasmic fractions, forming a complex that shuttles between nucleus and cytoplasm. Nuclear import of FAM98B requires active transcription, and silencing of hCLE downregulates nuclear and cytosolic accumulation of FAM98B. Nuclear/cytoplasmic fractionation, co-immunoprecipitation, photoactivatable GFP-hCLE live imaging, RNA silencing PloS one Medium 24608264
2019 FAM98B associates with hCLE (C14orf166), DDX1, and HSPC117 in a cap-binding complex; all four proteins bind cap analog-containing resins. This complex modulates mRNA translation, as hCLE silencing reduces accumulation of all four proteins and decreases mRNA translation. Cap analog affinity resin pulldown, competition/elution experiments, RNA silencing, polysome analysis Frontiers in physiology Medium 30833903
2016 FAM98B, along with its structural homolog FAM98A, forms a novel complex with DDX1 and C14orf166 that is required for PRMT1 expression in colorectal cancer cells. Knockdown of FAM98B suppresses cellular proliferation and colony formation. Knockdown of both FAM98A and FAM98B together does not cause additional reduction compared to single knockdowns, indicating they function in the same pathway axis. Co-immunoprecipitation, shRNA knockdown, cell proliferation and colony formation assays, epistasis by double knockdown The international journal of biochemistry & cell biology Medium 28040436
2021 The core of the human tRNA-LC is assembled from RTCB and the C-terminal alpha-helical regions of DDX1, CGI-99, and FAM98B, all of which are required for complex integrity. CGI-99 and FAM98B associate via their N-terminal domains to form a stable subcomplex. Crystal structure of GMP-bound RTCB reveals divalent metal coordination in the active site. Biochemical analysis of inter-subunit interactions, crystal structure determination of RTCB and CGI-99 N-terminal domain, deletion mutagenesis eLife High 34854379
2025 FAM98B contains the most glycine-rich intrinsically disordered region (IDR) in the human proteome. In GGC repeat expansion disorders, polyglycine aggregates sequester and deplete FAM98B through this glycine-rich IDR, disrupting tRNA-LC function and tRNA processing. Fam98b depletion in adult mice causes progressive motor coordination deficits and hindbrain pathology. Biochemical aggregation assays, patient tissue analysis, FISH for tRNA splicing intermediates, in vivo mouse knockdown with behavioral and histological readouts Science (New York, N.Y.) High 40674500
2020 FAM98B binds to the major allele of the rs11466313 variant in the 5' regulatory region of TGFB1; knockdown of FAM98B attenuates the enhanced TGFB1 promoter activity driven by the major allele, indicating FAM98B functions as a transcriptional activator of TGFB1 through this regulatory element. DNA pull-down assay, mass spectrometry identification, luciferase reporter assay, siRNA knockdown Breast cancer research and treatment Medium 32757134
2025 FAM98B depletion by shRNA in osteoclast precursors specifically disrupts lysosome trafficking and bone resorption in osteoclasts, with phenotypes similar to FAM98A knockdown. In Fam98a-null osteoclasts, increased Fam98b expression occurs as compensation. shRNA knockdown in osteoclast precursors, lysosomal trafficking assays, bone resorption assays, qPCR for compensatory expression Biology Medium 39857276
2025 Cryo-EM structure of the human tRNA-LC reveals that CGI-99, DDX1, and FAM98B form an alpha-helical bundle contacting RTCB. FAM98B and CGI-99 form an intricately co-folded heterodimer that clamps Ashwin in a pincer-like structure. FAM98A and FAM98C can substitute for FAM98B to underpin compositionally distinct RTCB-containing complexes that lack Ashwin and may have distinct cellular functions. Cryo-EM structure determination at atomic resolution, structure-based mutagenesis, interaction analysis bioRxivpreprint High bio_10.1101_2025.08.01.668197
2025 Three forms of the tRNA-LC exist depending on which FAM98 paralog (FAM98A, FAM98B, or FAM98C) is incorporated. Ashwin interacts exclusively with the FAM98B-containing complex, enabling nuclear localization of this specific tRNA-LC form for tRNA biogenesis. The FAM98B-tRNA-LC is thus the nuclear form required for pre-tRNA splicing, while FAM98A/C-containing complexes remain cytoplasmic for XBP1 mRNA splicing during UPR. Co-immunoprecipitation, NLS disruption mutagenesis, subcellular fractionation, tRNA splicing intermediate accumulation assay, rescue experiments with RTCB-NLS fusion bioRxivpreprint High bio_10.1101_2025.08.01.668163
2025 Cryo-EM structure of the five-subunit Danio rerio tRNA ligase complex shows that the DDX1 helicase module is mobile and can modulate the activity of RTCB. Chemical crosslinking confirmed specific RTCB residue involvement in RNA binding. Cryo-EM structure determination, thiol-based chemical crosslinking The Journal of biological chemistry Medium 40220997

Source papers

Stage 0 corpus · 14 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2020 Long non-coding RNAs (lncRNAs) in spermatogenesis and male infertility. Reproductive biology and endocrinology : RB&E 117 33126901
2014 Analysis of orthologous groups reveals archease and DDX1 as tRNA splicing factors. Nature 101 24870230
2014 hCLE/C14orf166 associates with DDX1-HSPC117-FAM98B in a novel transcription-dependent shuttling RNA-transporting complex. PloS one 52 24608264
2021 Molecular architecture of the human tRNA ligase complex. eLife 33 34854379
2016 FAM98A associates with DDX1-C14orf166-FAM98B in a novel complex involved in colorectal cancer progression. The international journal of biochemistry & cell biology 33 28040436
2019 hCLE/RTRAF-HSPC117-DDX1-FAM98B: A New Cap-Binding Complex That Activates mRNA Translation. Frontiers in physiology 20 30833903
2023 LncRNA-Gm9866 promotes liver fibrosis by activating TGFβ/Smad signaling via targeting Fam98b. Journal of translational medicine 9 37919785
2022 Joint Secondary Transcriptomic Analysis of Non-Hodgkin's B-Cell Lymphomas Predicts Reliance on Pathways Associated with the Extracellular Matrix and Robust Diagnostic Biomarkers. Journal of bioinformatics and systems biology : Open access 9 36873459
2025 Polyglycine-mediated aggregation of FAM98B disrupts tRNA processing in GGC repeat disorders. Science (New York, N.Y.) 8 40674500
2021 Role of C14orf166 in viral infection and RNA metabolism and its relationship with cancer (Review). Molecular medicine reports 3 33786620
2020 Effect of functional variant rs11466313 on breast cancer susceptibility and TGFB1 promoter activity. Breast cancer research and treatment 3 32757134
2025 FAM98 Family Proteins Play Distinct Roles in Osteoclastogenesis and Bone Resorption. Biology 1 39857276
2025 Structural and biochemical characterization of the 3'-5' tRNA splicing ligases. The Journal of biological chemistry 1 40220997
2007 [Gene screening in five Chinese families with hereditary spastic paraplegia with thin corpus callosum]. Zhonghua yi xue yi chuan xue za zhi = Zhonghua yixue yichuanxue zazhi = Chinese journal of medical genetics 1 18067082

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