DOLK catalyses the CTP-dependent phosphorylation of dolichol to dolichyl monophosphate
(Dol-P), the terminal step of de novo Dol-P biosynthesis at the ER membrane. Dol-P is the
essential lipid carrier that primes dolichol-linked oligosaccharide (LLO) assembly for
protein N-glycosylation and is also required for O-mannosylation, C-mannosylation and
GPI-anchor biosynthesis.
IntAct GO:0005515 protein binding annotations derive from large-scale binary interactome /
AP-MS maps (PMID:25416956, 26871637, 28514442, 32296183, 33961781). Interactors are largely
co-ER membrane proteins (ion channels KCNA1/3/6/10, CD79A, GPX8, EDA, etc.). None establish a
specific molecular function for DOLK; per curation policy protein binding is uninformative
and these are MARK_AS_OVER_ANNOTATED (not REMOVE — experimental IPIs).
This entry supersedes the earlier blanket interpretation of interaction screens, nucleolar staining and the duplicate glycan-supply core; the earlier notes remain intact. The 25 historical imported assertions retain all source fields. All 45 current GOA contexts map to those same 25 term/evidence/reference triples, with additional partner detail rather than new biological assertions. The inherited authored GO:0006488 proposal remains visible with retired: true; no imported GOA assertion is retired.
The single core is CTP-dependent dolichol phosphorylation at the ER membrane, directly producing Dol-P. The human cDNA and topology papers support this mechanism at their available complete-abstract scope. Later human patient-cell studies directly assay the reaction: PMID:17273964, Methods and Figure 4 uses crude fibroblast extracts, radiolabeled CTP, dolichol and a no-dolichol background; PMID:22242004(https://pubmed.ncbi.nlm.nih.gov/22242004/) independently measures Dol-P formation and uses sequence-verified human ORFs for yeast complementation. These assays are not purified-enzyme kinetics.
GO:0006488 describes glycosyl-chain assembly on the lipid carrier. DOLK supplies Dol-P, while downstream enzymes add the glycosyl residues. Rescue of CPY glycosylation and reduced LLO abundance establish downstream dependence, without identifying a DOLK glycosyl-addition step. Same-role comparator annotations are mixed: SRD5A3 carries GO:0006488, whereas the checked DHDDS, DOLPP1 and yeast SEC59 records do not. This is not evidence for a universal exclusion convention or a missing-annotation gap. The target GO-CAM models the established kinase/Dol-P process. The conservative revision withdraws the historical additional authored assertion rather than treating necessity alone as direct participation.
The positive EDA binary pair supports cytokine-partner binding, and the KCNA3 53–575 pair supports transmembrane-transporter binding. Neither asserts receptor agonism, channel regulation or a native DOLK signaling pathway. Three heterogeneous BioPlex/HuRI aggregate rows remain non-core. The executed interaction audit preserves all 42 raw fields, roles, regions, source contexts and independently computed sequence checks. Different actions for repeated partners reflect the scope of each historical aggregate. No interaction is rejected merely because its term is generic.
PMID:26871637, Figure 6D and Methods reports divergent yeast rescue and interaction profiles for historical clones DOLK_1 and DOLK_2: DOLK_1 rescues and binds KCNA3, whereas DOLK_2 does not rescue and binds UBQLN1 splice constructs. The figure itself was inspected. Clone-sequence supplements were not recovered, so these names are not assigned to additional official isoforms. The current official record contains one 538-residue protein and no alternative-products block. A failed yeast rescue in that construct context is not universal inactivity in every setting.
The target-resolved HPA XML supports nucleolar staining with two approved ICC/IF antibodies, including nucleolar observations in three cell lines for HPA068706. Multipass ER topology alone does not establish that this signal is background. The observation is retained as contextual localization, without assigning nucleolar kinase activity or claiming that image pixels or genetic antibody controls were assessed.
PAINT tree, alignment and the three literal IBD placements support the target lineage and the established activity/process/location. Target experimental evidence within the IBD descendants is legitimate. Reactome's normal reaction and the failed-reaction variant set are treated separately; a gene-level kinase assignment does not imply normal activity for every pathogenic variant.
The source and assay guide supplies public source locators, model paths and access limits. Existing publication files and flags are unchanged. The configured Falcon research attempt failed with HTTP 403 and its configured fallback failed with HTTP 401; no provider report was fabricated. Manual evidence and notes supply the assessment.
The binding decisions retain the positive results while separating partner identity from native physiological function. The current cytokine-binding definition does not require receptor activity or an extracellular module in the binding protein. The EDA reference identifies both membrane and secreted forms of the cytokine; the records do not establish which processed species or surface made contact. The positive 2014 validated pair and HuRI validated pair have the same bounded class interpretation. The broader HuRI row also retains its other positive pairings; its non-core action is not a negative judgment on EDA or KCNA3. The raw array EDA record with a DOLK residue-441 perturbation is distinguished from the validated record with no such feature.
For KCNA3, Figure 6D and Results directly distinguish DOLK_1 from DOLK_2. The validated record identifies KCNA3 residues 53–575. The transporter-binding refinement remains restricted to that positive clone result; an unqualified gene-level annotation is not proof that the current canonical sequence or all splice forms were tested. Historical clone sequences remain unresolved. The published interaction audit and its sequence-reference checks are unchanged.
The positive HPA nucleolar evidence is stated explicitly in the row reason and source guide. It is retained without inferring a nuclear catalytic role. The description restores the enzyme family and EC identifier and separates carrier production from the downstream uses of Dol-P and Dol-P-mannose. The existing 16923818 and 22242004 anchors are preserved; one short 12213788 microsomal-activity anchor is added only to the core. Existing publication access flags and separately recovered original scopes are unchanged.
The exact-head ClinGen instruction explicitly retains supported generic binding as non-core when a more informative activity is not established for the retained association scope. It takes precedence for this project over the general informational-exclusion recommendation. Published validation passed with three protein-binding advisories on the aggregate rows; this is an acknowledged task-specific exception, not a claim that the repository-wide rule changed. The mixed SRD5A3 comparator and retirement of the inherited authored GO:0006488 proposal remain unchanged.