MPK6 (Q39026, At2g43790) — curation notes
Identity and core molecular function
- Arabidopsis mitogen-activated protein kinase 6; CMGC/MAPK family, TEY-motif (TXY motif at 221-223). EC 2.7.11.24. Kinase domain residues 63-348; ATP-binding K92 (invariant Lys) essential.
- Core MF = MAP kinase activity (GO:0004707) / protein serine/threonine kinase activity. Activated by dual phosphorylation on Thr221/Tyr223 PMID:10713056.
- Substrate consensus is proline-directed L/P-x-S/T-P-R/K PMID:22631074.
- K92M/K93M abolish kinase activity (UniProt MUTAGEN) PMID:11875555. Crystal structures 5CI6, 6DTL.
Upstream activators (MAPKKs) — these are the IPI "protein binding" partners
- MKK2 (Q9S7U9), MKK4 (O80397), MKK5 (Q8RXG3), MKK6 (Q9FJV0); also MKK3, MKK7, MKK9. MAPKKKs: MEKK1, YODA, ANP1, MKKK20.
- These IPI bare protein-binding annotations (PMID:15225555, 19513235, 21798944, 32612234, 27913741) are uninformative and were marked REMOVE; the cascade relationship is captured by MAPK cascade (GO:0000165).
Substrates (also annotated as bare protein binding -> REMOVE)
- PP2C AP2C1; PTP1/MKP1 (O82656) [PMID:19789277, PMID:27029354]; MKP2/DSPTP1B (Q9M8K7) PMID:20626661.
- PTP1 IPI from PMID:27029354 is annotated as phosphatase binding (GO:0019902) — kept as ACCEPT since it is more informative than bare protein binding.
Localization
- Cytoplasm (resting) and nucleus (upon activation/phosphorylation) [PMID:15500467 "activation and nuclear translocation ... during ozone exposure"; PMID:31235876 nuclear, stomatal].
- Cytosol HDA PMID:28887381. Cell cortex via BASL recruitment PMID:25843888. Preprophase band, phragmoplast, TGN, PM in dividing root cells PMID:19832943.
Pleiotropy / biological processes (mostly KEEP_AS_NON_CORE)
- Immunity (FLS2->MEKK1-MKK4/MKK5-MPK3/MPK6) PMID:11875555; camalexin biosynthesis PMID:18378893; priming PMID:19318610; bacterial/fungal defense [PMID:11875555, 21947882].
- Hormones: ethylene PMID:12628921, jasmonate (MKK3-MPK6, MYC2) PMID:17369371, ABA (MKK1/MKK5) [PMID:18248592, 27913741].
- Abiotic stress: ROS/oxidative/H2O2 [PMID:10717008, 11577197, 17933903], osmotic PMID:12220631, cold/salt (MKK2) PMID:15225555, freezing (ICE1) PMID:29056553, UV-B PMID:21790814, abiotic activation PMID:11123804, harpin PMID:11500556.
- Development: stomatal patterning/ACD [PMID:25843888, 31235876], inflorescence architecture (ERECTA) PMID:23263767, ovule PMID:18364464, pollen development/tube guidance [PMID:24830428, 24717717], root development/cell division PMID:19832943, leaf senescence (MKK9) PMID:19251906, L-glutamate root response PMID:29344832.
- Strong functional redundancy with MPK3 throughout (mpk3 mpk6 double often embryo-lethal).
Curation decisions summary
- 76 existing_annotations reviewed (matches 76 non-header GOA rows after header).
- ACCEPT (28): core MF (MAP kinase / protein serine kinase / protein kinase activity, ATP binding), MAPK cascade, signal-transduction terms, principal localizations (nucleus, cytoplasm, cytosol), phosphatase binding, protein phosphorylation.
- KEEP_AS_NON_CORE (33): the many downstream biological processes and specialized localizations (cell cortex, TGN, phragmoplast, preprophase band) for this pleiotropic kinase.
- REMOVE (15): all 15 bare GO:0005515 protein binding IPI entries (uninformative; partners are MKKs/substrates better captured by cascade/kinase terms). Phosphatase binding (GO:0019902) retained as the informative alternative for phosphatase interactions.
- No publications were inaccessible (all 43 cached), so no UNDECIDED was needed.
Proposed
- New MF term idea: "MAP kinase substrate binding" (child of protein kinase binding GO:0019901) to capture the many kinase-substrate IPIs informatively instead of discarding them as generic protein binding.
Deep research synthesis (Falcon / Edison Scientific, file:ARATH/MPK6/MPK6-deep-research-falcon.md)
The Falcon deep-research report corroborates and does not contradict the existing review. Key points used to strengthen supported_by evidence:
- Core MF: MPK6 is a "Proline-directed CMGC MAP kinase; terminal kinase in MAPK cascades activated by dual Thr/Tyr phosphorylation in the TxY motif" that "phosphorylates downstream substrates including transcription factors, enzymes, RNA-metabolism proteins, and developmental regulators, often redundantly with MPK3." Added to MAP kinase activity, protein serine kinase activity and core_functions.
- MAPK cascade architecture: "MAPKs are terminal kinases in a conserved signaling architecture in which upstream kinases sequentially activate MAP2Ks and then MAPKs." Added to GO:0000165.
- Immunity (defense response to bacterium): "the MEKK1–MKK4/5–MPK3/MPK6 PTI module downstream of FLS2 that contributes to pathogen resistance"; "MPK6 is a core component of immune MAPK cascades activated by pattern recognition receptors."
- Ethylene: "MPK6 phosphorylates ACC synthase isoforms (e.g., ACS6) to affect stability and ethylene output" — supports response to ethylene.
- Development (stomatal): "MPK3/MPK6 phosphorylate SPEECHLESS (SPCH) downstream of the YODA–MKK4/5 module, modulating stomatal initiation and lineage progression" — supports nucleus localization and developmental core function.
- Freezing/ICE1: compiled substrates "include MYB41, MYB15, HsfA2, ICE1, DCP1 ... with outcomes such as enhanced DNA binding, altered protein stability, and nuclear accumulation" — supports response to freezing (ICE1).
- Localization: substrates span nucleus and cytoplasm — "This implies a functional distribution spanning nucleus and cytoplasm"; "classic reports note stress-triggered nuclear translocation of MAPKs."
- New 2023-2024 findings NOT added as GOA annotations (no verifiable GO IDs in current GOA/UniProt): MPK6 in stress granules / P-bodies via DCP1 phosphorylation (He et al. 2024 preprint); MYB36 phosphosites in endodermal SGN3/CIF signaling (Ma et al. 2024, Nature Plants); cat2-1 / glutathione modulation of flg22-induced MPK3/6 kinetics (Yang et al. 2024 preprint). These are recorded here for future review but are largely preprints and would need verifiable curated GO terms before annotation.
- No action: UNDECIDED entries existed; none required resolution (all 43 cited publications were accessible).
PR #1417 review fix (2026-06-06): title-as-evidence supporting_text
- GO:0019902 phosphatase binding (original_reference_id PMID:27029354, IPI vs PTP1/O82656 — confirmed against GOA line 55). The original reference IS correct: PMID:27029354 directly documents the MPK6-PTP1 physical interaction by BiFC ("PTP1 interacted with MPK6 in the cytoplasm and nucleus (Fig. 7, lane 4, panel 4)"). The problem was the supporting_text being the bare paper title (about SnRK1). Replaced with that verbatim BiFC quote and added PMID:19789277 via additional_reference_ids with verbatim quote ("PROTEIN TYROSINE PHOSPHATASE1 (PTP1) also interacts with MPK6, but the ptp1 null mutant shows no aberrant growth phenotype"). original_reference_id left as PMID:27029354 per GOA.
- Other bare-title supporting_text replaced with real verbatim quotes on ACCEPT/KEEP annotations:
- GO:0005634 nucleus (IEA, PMID:15500467) and GO:0005737 cytoplasm (PMID:15500467): title fragment -> "activated AtMPK3 and AtMPK6 are translocated to the nucleus during the early stages of O3 treatment".
- GO:0005938 cell cortex (two entries, PMID:25843888): title / paraphrase -> verbatim "Phosphorylated BASL functions as a scaffold and recruits the MAPKKK YODA and MPK3/6 to spatially concentrate signaling at the cortex". (The earlier "Recruited by BASL at the cell cortex in a polarized manner" was a paraphrase, not an exact substring.)
- Remaining bare-title supporting_text instances are all on action: REMOVE annotations (uninformative protein binding being removed) and high-throughput interactome screens; left as-is as lower priority.
- Validation:
uv run ai-gene-review validate genes/ARATH/MPK6/MPK6-ai-review.yaml -> Valid (2 pre-existing unrelated warnings). All supporting_text now exact substrings of cited sources.
PR review follow-up: title-as-supporting_text on non-REMOVE annotations (2026-06-07)
Reviewer (ai4c-agent) flagged ~19 annotations whose supporting_text was merely the cited paper's title. Focused on the NON-REMOVE subset (ACCEPT / KEEP_AS_NON_CORE). Replaced bare titles with verbatim excerpts:
- GO:0106310 protein serine kinase activity (ACCEPT, PMID:15500467) -> "transient activation of 43 and 45 kDa MAPKs. These were identified as AtMPK3 and AtMPK6".
- GO:0009738 ABA-activated signaling (KEEP, PMID:27913741) -> "The activity of MPK6 was increased by ABA ... the AIK1-MKK5-MPK6 cascade functions in the ABA regulation of primary root growth and stomatal response".
- GO:1902065 response to L-glutamate (KEEP, PMID:29344832) -> "In-gel phosphorylation assays revealed a rapid and dose-dependent induction of MPK6 and MPK3 activities ... in response to L-Glu".
- GO:0006468 protein phosphorylation (ACCEPT, PMID:29056553) -> "mitogen-activated protein kinase 3 (MPK3) and MPK6 interact with and phosphorylate ICE1, which reduces its stability and transcriptional activity".
- GO:0050826 response to freezing (KEEP, PMID:29056553) -> "the mpk3 and mpk6 single mutants and the mpk3 mpk6 double mutants show enhanced freezing tolerance, whereas MPK3/MPK6 activation attenuates freezing tolerance".
- GO:0009555 pollen development (KEEP, PMID:24830428) -> "epitope-tagged WRKY34 is temporally phosphorylated by MPK3 and MPK6 ... at early stages in pollen development".
- GO:0009620 response to fungus (KEEP, PMID:21947882) -> "It was observed that the expression of both MAP2K9 and MAPK6 simultaneously increased up to middle stage of disease progression".
- GO:0010224 response to UV-B (KEEP, PMID:21790814) -> "The MKP1-interacting proteins MPK3 and MPK6 are activated by UV-B stress ... mutants mpk3 and mpk6 exhibit elevated UV-B tolerance".
- GO:0080136 priming of cellular response to stress (KEEP, PMID:19318610) -> "priming is associated with accumulation of mRNA and inactive proteins of ... MPK3 and MPK6. Upon challenge exposure ... more strongly activated in primed plants".
- GO:0048481 plant ovule development (KEEP, PMID:18364464) -> "two closely related mitogen-activated protein kinases ... MPK3 and MPK6, share a novel function in ovule development ...".
- GO:0042542 response to hydrogen peroxide (KEEP, PMID:17933903) -> "MPK6 and MPK7 were both activated by H(2)O(2), but only MPK7 activation was enhanced by MKK3".
Set to UNDECIDED (no quotable evidence in cached source):
- GO:0005829 cytosol (was ACCEPT, HDA, PMID:28887381): cached publication text (a global membrane-protein oligomerization proteomics study) does not mention MPK6/MAPK6 or its localization; the assignment lives only in supplementary dataset tables not present in the cached markdown, so no verbatim excerpt can substantiate it. Action -> UNDECIDED.
REMOVE annotations with bare-title supporting_text left as-is (out of scope; REMOVE stands on the protein-binding curation guideline).