2026-09-28. Initial assessment of the source annotations and primary literature.
The initial review has 19 source assertions, no NOT assertions or annotation-specific isoform flags, and two recorded products: Q8IZT6-1 and Q8IZT6-2 (VSP_010680). The UniProt sequence is 3477 aa; isoform 2 lacks residues 1356–2940. These records are preserved. Older paper motif counts depend on prediction conventions and do not replace current sequence features with the paper's 81-versus-14 count.
The original PMID set is 15972725 and 21044324. The first cache is abstract-only; the second contains XML-extracted full text. The XML cache repeats nested sections; the substantive Results, Discussion and Methods paragraphs were read separately, including main and supplementary figure legends. Figure pixels and supplementary movies were not inspected.
Human HeLa, U2OS, HDF and SH-SY5Y experiments are distinguished from monkey COS-7 work. Three antibodies gave similar staining; only the N-terminal 217-2 antibody was optimized for endogenous immunoblotting. Peptide/preimmune specificity controls are reported, some data not shown. Interphase ASPM is predominantly nuclear, and this study did not detect an interphase centrosomal pool. After nuclear-envelope breakdown it forms a pole-proximal ring around, rather than extensively overlapping, gamma-tubulin. Late mitotic protein localizes at central-spindle minus ends and a narrow midbody ring.
Nocodazole removes spindle-associated ASPM while retaining gamma-tubulin; taxol produces ASPM-positive aster centers and separate ASPM-negative anastral gamma-tubulin foci. This supports microtubule-dependent localization and should not be converted into a constitutive centriolar-core claim.
Two independent U2OS siRNAs (ASP1 and ASP2) reduce spindle-pole ASPM, with different efficacy. The paper did not detect a matching decrease of total interphase protein. Live-cell orientation/cytokinesis analysis used ASP1 and GL3 control, pooling four experiments (94 and 126 divisions). The paper's word 'asymmetric' denotes geometric division orientation relative to the dish, not an independently measured daughter-cell fate. The cytokinesis deficit exceeds the effect of altered orientation alone. No new apoptosis-regulation process is inferred from downstream caspase activation.
Patient fibroblasts with IVS25+1G>T retain an apparently stable protein but show markedly reduced pole localization; cDNA reveals a cryptic splice event deleting a tripeptide at the C terminus. The body and one figure legend disagree about upstream/downstream wording; the sequenced body says upstream. Keep current canonical protein length rather than repeating the isolated 3447-aa typo. Human C-terminal D1–D3 fragments (3177–3477, 3256–3477 and 3315–3477) expressed in HeLa and COS-7 cause dominant-negative spindle/cytokinesis defects. These are fragments, not evidence that canonical isoform 2 has all the same effects.
The spindle-organization and spindle-positioning processes are supported by actual structural localization plus perturbations. The discussion offers several possible late-cytokinesis mechanisms; it does not establish a direct motor, actin-binding enzyme or cytokinesis kinase function for ASPM. Later PMID:28883092 provides a useful compensation context rather than a reason to erase these experimental annotations.
Original PLOS article: complete Results through Figure 6 and targeted Methods read. Native mouse oocytes and mouse embryonic fibroblasts were studied. ASPM decorates the meiotic spindle and co-immunoprecipitates with calmodulin; the immunoprecipitation control is unrelated rabbit IgG. This is cellular association, not an isolated purified binary-binding assay. One ASPM morpholino reduces the measured protein signal and disrupts spindle morphology/progression; uninjected and control-morpholino controls are included. No rescue experiment is claimed. Spindle-checkpoint machinery is discussed as a hypothesis, not measured directly. These data support the mouse donor biology for existing human orthology transfers; they are not direct human oocyte assays.
The official NCBI rat Aspm record links this PMID to its IDA apical-membrane, centrosome, midbody and mitotic-pole rows. Thus the rat donor underlying the human IEA is identifiable, not a guessed citation.
Direct publisher and author PDF opening returned 403. Indexed original Methods and Figure 1/3 legends were readable at the author-hosted manuscript and publisher PDF. Methods specify rat E14-brain cDNA for ASPM MTB/CTR constructs and a rat N-terminal antigen for Ab418; BL2048 is raised against a human peptide. HeLa/HEK293 expression hosts are human, whereas these expressed ASPM fragments are rat. Figure 1 demonstrates endogenous HeLa prophase centrosomal, metaphase spindle-pole and cytokinetic midbody staining with two antibodies. Figure 3 explicitly shows E13 rat neocortical ASPM at neuronal-progenitor poles and the ventricular apical surface, with aurora-B association at that surface. This is sufficient to avoid declaring the rat source nonexistent, but full Results, complete controls and pixels have not been read. The non-core human apical annotation remains a candidate for cautious retention or UNDECIDED after consultation, not automatic removal because it is unusual.
The official GO term page was read via its indexed content: GO:0051011 is microtubule minus-end binding, a molecular function and child of GO:0008017 microtubule binding. QuickGO returned only a JavaScript shell/API failure; there is no callable OLS tool in this session.
PMID:28436967(https://pmc.ncbi.nlm.nih.gov/articles/PMC5458804/) supports a conserved minus-end mechanism, but the purified reconstitution uses short mouse ASPM plus calmodulin. This species boundary was independently checked. Endogenous human ASPM is recruited to newly generated minus ends in the cellular experiment. The mouse purified assay is not a human IDA, and katanin supplies ATP-dependent severase activity. At this initial reading, a new human binding annotation remained under consideration; the final inference is recorded below.
Additional literature identified at the initial reading comprised PMIDs 28436967, 27562601, 28883092, 23152892 and 17534152. PMID:19219036 remains separate worm comparator evidence.
All 19 original source assertions and both alternative products are preserved. Decisions are 12 ACCEPT, 6 KEEP_AS_NON_CORE and 1 UNDECIDED, plus one NEW ISS molecular-function inference for microtubule minus-end binding. The human cellular observations and mouse ASPM-calmodulin reconstitution are distinguished. Katanin supplies severing catalysis; no new process, motor, ATPase, kinase or human-isoform-specific activity is proposed. Calmodulin association supports the integrated microtubule mechanism without requiring a separate core-function entry. The unresolved apical-membrane assignment retains positive rat evidence without claiming that midbody staining establishes a membrane domain.
The normal deep-research attempt failed and generated no provider report. The two seed publications and five additional sources were recovered by normal fetchers before final authoring. Cached full-text availability is recorded separately from actual scientific reading scope.
The earlier final-synthesis paragraph records the initial publication. This follow-up retains all 19 original source assertions, the existing NEW ISS minus-end-binding assertion, both products and one core. The only action change is row 14 (zero-based): spindle localization becomes MODIFY with establishment of mitotic spindle orientation. Current decisions are 11 ACCEPT, 6 KEEP_AS_NON_CORE, 1 MODIFY, 1 UNDECIDED and 1 NEW. All original nine literal supporting quotations and cached source bytes are preserved.
The official GO:0000132 record defines mitotic spindle alignment relative to other cellular structures. Its parents include GO:0051294 establishment of spindle orientation and GO:0040001 establishment of mitotic spindle localization; the displayed ancestry reaches the original GO:0051653 spindle localization. The term is not human-specific: the human restriction belongs to the experiments. The original U2OS study measures alignment relative to the dish, and the human knockout/interface experiments in PMID:28436967(https://pmc.ncbi.nlm.nih.gov/articles/PMC5458804/) corroborate mitotic misorientation. This is a refinement of the source assertion, not a second NEW process row.
The core now names ASPM's active outputs. Actual normal XML Results in PMID:28436967 show that purified short mouse ASPM with calmodulin slows minus-end growth and blocks outgrowth at higher concentrations, and that ASPM recruits katanin and enhances its severing activity. Mutation-based recruitment experiments separate this assistance from the severase chemistry supplied by katanin. The statement that other MAPs were not detected supports autonomous ASPM end tracking in that preparation; it does not convert the preparation into isolated human ASPM. Endogenous tagged human ASPM co-purifies with calmodulin and katanin and is recruited to newly exposed minus ends. Those human observations support the conserved mechanism without directly measuring isolated human minus-end growth inhibition. The complete primary Results paragraphs for these targeted experiments were reread; figure pixels, movies, raw spectra and complete supplementary datasets remain unread.
GO:0031115 denotes reduced microtubule polymerization; its parents are regulation terms, with an explicit negatively_regulates link to GO:0046785 microtubule polymerization. It is not an orientation ancestor or a synonym for depolymerization. The purified mouse growth-blocking experiment meets participation directly, not merely through necessity. Nevertheless, this bounded human follow-up retains the existing conserved binding inference and makes its outputs explicit in prose without adding a second NEW process assertion or another core BP. This is a conservative scope and species-inference boundary, not a claim that ASPM is passive or that ortholog inference is invalid.
For PMID:27562601(https://pmc.ncbi.nlm.nih.gov/articles/PMC5048379/), the complete normal abstract and indexed original ASPM/CITK Results, Figure 4 legend and adjacent rescue/paclitaxel Results, plus cell-culture and transfection/RNAi/rescue Methods were read. ASPM-depleted human HeLa cells lose astral microtubules and spindle alignment; CITK overexpression and low-dose paclitaxel rescue the orientation phenotype. The CITK rescue construct is mouse, and the later EB3 dynamics experiments primarily interrogate CITK. These are targeted human-cell data supporting the orientation refinement, not an isolated ASPM polymerase assay. The normal cache remains abstract-only and its availability flags remain accurate; external indexed reading is recorded separately.
The cytokinesis question is explicit. In PMID:21044324, failures also occur in geometrically symmetric divisions, and asymmetric division does not fully account for the excess failures. Thus cytokinesis is not dismissed as simply caused by misorientation. The Discussion presents alternative executing mechanisms involving central-spindle organization, astral microtubules or coupling to the cortex, without resolving which late-cytokinesis step ASPM directly performs. No NEW cytokinesis BP follows solely from depletion and localization.
The apical-plasma-membrane row remains UNDECIDED. Positive rat apical-surface/midbody-ring evidence is retained; incomplete indexed-source access does not establish that the original membrane assignment was an over-annotation. The standalone description now includes autosomal recessive primary microcephaly type 5 (MCPH5), as recorded in the preserved human UniProt entry, and keeps experimental provenance in reasons and notes.