Extended Data Fig. 2: Amino acid sequence characteristics of Trx-h2 and its specific interaction with CBF1 at low temperature.

a, Comparison of N-terminal amino acid sequences (~80 residues) of 11 cytoplasmic Trx-hs in Arabidopsis, and their putative acylation sites predicted by the TermiNator program. Based on the modification pattern of fatty acids, yellow, magenta, green, and cyan boxes on the left represent subgroup I, Sub-II, Sub-III, and Sub-IV Trx-hs, respectively. Critical amino acid residues required for the myristoylation, N-α-acetylation, and palmitoylation of Trx-hs are outlined by a red box (Gly2 of Sub-II), blue box (Ala2 of Sub-I), and green box (Cys5 of Sub-III & IV), respectively. The active site Cys residues (CXXC motif) are outlined by a maroon box. b, Schematic representation of Trx-h2 and its point mutation variants, Trx-h2(G/A) and Trx-h2(C/S). c, Sequence features of Trx-h2. Active site Cys residues of Trx-h2 (at amino acid positions 59 and 62) are indicated in bold blue font, and the Gly2 residue required for myristoylation (G²) is indicated in green. The conserved Trx motif (122 amino acids) is indicated in red. The bipartite nuclear localization signal (NLS) sequence identified from the NLS-mapper program¹⁸ is underlined. The asterisk at the end of the protein sequence indicates the stop codon. d, Interacted specificity of CBF1 with Trx-h2, but not with Trx-h3 (control), analyzed by BiFC at 4 °C. Scale bars = 20 μm.