Figure 1.

Characterization of NUA T-DNA Insertion Lines.

(A) The coiled-coil domain prediction of NUA (At1g79280) using COILS, and comparison with human Tpr, Drosophila Megator, and yeast Mlp1 and Mlp2. Black bars show predicted coiled-coil domains. The dimerization and NPC association domain of Tpr are also depicted. Bar = 200 amino acids (aa).

(B) Position of T-DNA insertions and PCR primers. The confirmed insertion sites in nua-1, nua-2, nua-3, and nua-4 are indicated by vertical arrows above and below the schematic exon-intron structure of the NUA gene. Exons are depicted as gray bars and introns as black lines. Horizontal arrows indicate the positions of PCR primers, and brackets indicate the positions of the RT-PCR products shown in (D). The area surrounding the nua-2 insertion site is shown enlarged, and the structure of the ∼200 and ∼300-bp fragments derived from RT-PCR with NUA-D primers in nua-2 is indicated. ex, exon; int, intron.

(C) Immunoprecipitation of NUA from Arabidopsis wild-type and nua-1, nua-2, and nua-3 seedlings. Arrowheads depict NUA bands, and asterisks mark unspecific IgG bands also seen without plant extract (data not shown). Molecular weights are indicated on the left.

(D) RT-PCR products derived from primer pairs indicated on the right (as shown in [B]) and plant tissues as indicated on the top. Approximate sizes of the fragments amplified with the primer pair for NUA-D in nua-2 are indicated on the left. Two exposures are shown for the fragments amplified with NUA-D primers. The primer combinations in each reaction were as follows: NUA-A, F1 + LP115409; NUA-B, F1 + SALK_057101LP; NUA-C, RP057101.1 + R1(1599); NUA-D, RP069922 + LP069922; NUA-E, CS821281FP + R2(3798); NUA-F, RP079795 + R3(4993). All primer sequences are listed in Supplemental Table 2 online. Tub, tubulin 2 (At5g62690).