Abstract
Protein kinases are major signaling molecules that are involved in a variety of cellular processes. However, the molecular mechanisms whereby protein kinases discriminate specific substrates are still largely unknown. Ca(2+)-dependent protein kinases (CDPKs) play central roles in Ca(2+) signaling in plants. Previously, we found that a tobacco (Nicotiana tabacum) CDPK1 negatively regulated the transcription factor REPRESSION OF SHOOT GROWTH (RSG), which is involved in gibberellin feedback regulation. Here, we found that the variable N-terminal domain of CDPK1 is necessary for the recognition of RSG. A mutation (R10A) in the variable N-terminal domain of CDPK1 reduced both RSG binding and RSG phosphorylation while leaving kinase activity intact. Furthermore, the R10A mutation suppressed the in vivo function of CDPK1. The substitution of the variable N-terminal domain of an Arabidopsis thaliana CDPK, At CPK9, with that of Nt CDPK1 conferred RSG kinase activities. This chimeric CDPK behaved according to the identity of the variable N-terminal domain in transgenic plants. Our results open the possibility of engineering the substrate specificity of CDPK by manipulation of the variable N-terminal domain, enabling a rational rewiring of cellular signaling pathways.
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Claims 7
Claim · model-written summary
CDPK1 variable N-terminal domain is necessary for RSG recognition.
- Taxon
- tobacco
Nicotiana tabacume0 - Gene
- CDPK1e1
- Outcome
- recognition of RSGe1
- Qualifier
- variable N-terminal domaine1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“a tobacco (Nicotiana tabacum) CDPK1”
e1“the variable N-terminal domain of CDPK1 is necessary for the recognition of RSG.”
Claim · model-written summary
R10A reduced RSG binding.
- Taxon
- tobaccoNicotiana tabacume0
- Treatment
- mutation (R10A)e1
- Gene
- CDPK1e1
- Outcome
- RSG bindinge1
- Direction
- reducede1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“a tobacco (Nicotiana tabacum) CDPK1”
e1“A mutation (R10A) in the variable N-terminal domain of CDPK1 reduced both RSG binding and RSG phosphorylation while leaving kinase activity intact.”
Claim · model-written summary
R10A reduced RSG phosphorylation.
- Taxon
- tobaccoNicotiana tabacume0
- Treatment
- mutation (R10A)e1
- Gene
- CDPK1e1
- Outcome
- RSG phosphorylatione1
- Direction
- reducede1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“a tobacco (Nicotiana tabacum) CDPK1”
e1“A mutation (R10A) in the variable N-terminal domain of CDPK1 reduced both RSG binding and RSG phosphorylation while leaving kinase activity intact.”
Claim · model-written summary
R10A left kinase activity intact.
- Taxon
- tobaccoNicotiana tabacume0
- Treatment
- mutation (R10A)e1
- Outcome
- kinase activitye1
- Direction
- intacte1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“a tobacco (Nicotiana tabacum) CDPK1”
e1“A mutation (R10A) in the variable N-terminal domain of CDPK1 reduced both RSG binding and RSG phosphorylation while leaving kinase activity intact.”
Claim · model-written summary
R10A suppressed CDPK1 in vivo function.
- Taxon
- tobaccoNicotiana tabacume0
- Treatment
- R10A mutatione1
- Outcome
- in vivo function of CDPK1e1
- Direction
- suppressede1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“a tobacco (Nicotiana tabacum) CDPK1”
e1“the R10A mutation suppressed the in vivo function of CDPK1.”
Claim · model-written summary
Replacing the At CPK9 N-terminal domain with that of Nt CDPK1 conferred RSG kinase activity.
- Taxon
- Arabidopsis thaliana
Arabidopsis thalianae0 - Treatment
- Nt CDPK1e0
- Outcome
- RSG kinase activitiese0
- Direction
- conferrede0
- Comparator
- At CPK9e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Arabidopsis thaliana CDPK, At CPK9, with that of Nt CDPK1 conferred RSG kinase activities.”
Claim · model-written summary
Chimeric CDPK behavior in transgenic plants followed variable N-terminal domain identity.
- Taxon
- TobaccoNicotiana tabacumt0
- Outcome
- chimeric CDPK behavede0
- Qualifier
- identity of the variable N-terminal domaine0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“This chimeric CDPK behaved according to the identity of the variable N-terminal domain in transgenic plants.”
t0“Tobacco”
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See all 8 papers →Claims are grouped by organism, treatment and what was measured or targeted. A paper counts as positive when any of its claims in the group is positive.