Table 1.
Distribution of FLZ gene family in sequenced genomes.
Figure 1.
Alignment between FLZ and zf-FCS domain.
(A) Multiple sequence alignment between FLZ and zf-FCS domains. Conserved cysteine, phenyl alanine and serine residues are marked by asterisks. (B) HMM-HMM alignment between FLZ domain and zf-FCS showing similarity in sequence conservation.
Figure 2.
Secondary structure pattern and sequence conservation of FLZ domain.
(A) Secondary structure conservation of FLZ domain. Red color indicates alpha helix and blue color indicates beta-sheet. Confidence gradient of secondary structure formation is given on the top. (B) Sequence logo of Arabidopsis, Medicago, poplar and rice FLZ domains showing amino acid conservation.
Figure 3.
Schematic representation of domain organization in FLZ protein family.
The FLZ proteins are scanned by InterProScan to identify conserved domains. A. thaliana FLZ1 is shown as a representative model for proteins which contain FLZ domain only. Proteins which possess other domains along with FLZ are also shown. The domains are abbreviated as follows, FLZ (FCS like zinc finger, PF04570), PPR (Pentatricopeptide Repeat, PF01535), Cupin (Cupin 1, PF00190), ICF (Ion channel family, PF00520), CND (Cyclic nucleotide-binding domain, PF00027) and DUF3354 (Domain of unknown function 3354, PF11834).
Figure 4.
FLZ acts as the module for protein-protein interaction.
(A) FLZ1 interacts with PFA-DSP3 and STH2 in Y2H. Murine p53 and SV40 large T-antigen interaction taken as positive control and p53 and lamin interaction is taken as negative control. (B) The deletion constructs of FLZ1, N-terminal (1–88 amino acids), FLZ (89–140 amino acids) and C-terminal (141–177 amino acids). (C) Y2H with deletion constructs of FLZ1 with PFA-DSP3 and STH2 showing FLZ is essential for their interaction. (D) And (E) beta-galactosidase activity of full length FLZ1 and deletion constructs interaction with PFA-DSP3 and STH2.
Figure 5.
FLZ domain mediates the interaction of FLZ1 and PFA-DSP3.
(A) BiFC, FLZ1 and PFA-DSP3 interact exclusively in nucleolus. Upper panel: whole cell, YFP alone, DAPI alone and merged. Lower panel: nucleus zoomed, YFP alone, DAPI alone and merged. (B) BiFC of PFA-DSP3 with deletion constructs of FLZ1: N-terminal (1–88 amino acids), FLZ (89–140 amino acids) and C-terminal (141–177 amino acids). (C) BiFC of PFA-DSP3 with AT5G49120: YFP alone, DAPI alone and merged. YFP were excited at 514 nm and emission was recorded at 530 nm. DAPI were excited at 351 nm and emission was recorded at 450 nm.
Figure 6.
Sub-cellular localization of FLZ1 and PFA-DSP3 in onion epidermal cells.
(A) Vector alone. (B) FLZ1. (C) PFA-DSP3. Left to right, YFP only, bright field, merged. FLZ1 is localized in both nucleus and cytoplasm while PFA-DSP3 is exclusively localized in nucleus. YFP were excited at 514 nm and emission was recorded at 530 nm.