Results: 5

1.
F <span style="font-variant: small-caps" class="small-caps">igure</span>  3.—

F igure 3.—. From: Point Mutations in the Stem Region and the Fourth AAA Domain of Cytoplasmic Dynein Heavy Chain Partially Suppress the Phenotype of NUDF/LIS1 Loss in Aspergillus nidulans.

DAPI staining of various strains. All samples were from 42° cultures except for the two 32° samples that are specifically labeled. Bar, ∼5 μm.

Lei Zhuang, et al. Genetics. 2007 March;175(3):1185-1196.
2.
F <span style="font-variant: small-caps" class="small-caps">igure</span>  1.—

F igure 1.—. From: Point Mutations in the Stem Region and the Fourth AAA Domain of Cytoplasmic Dynein Heavy Chain Partially Suppress the Phenotype of NUDF/LIS1 Loss in Aspergillus nidulans.

Positions of the two nudF suppressor mutations in the dynein heavy chain (nudA) of A. nidulans. (A) The domain organization of the dynein heavy chain. Positions of the nudAL1098F and the nudAR3086C mutations are indicated by red arrows. (B) Sequence alignments of the regions surrounding the nudAL1098F and the nudAR3086C mutations, respectively. Accession numbers for human, A. nidulans, N. crassa, and S. cerevisiae dynein heavy chains are Q14204, XP_657722, EAA33380, and NP_012980, respectively.

Lei Zhuang, et al. Genetics. 2007 March;175(3):1185-1196.
3.
F <span style="font-variant: small-caps" class="small-caps">igure</span>  4.—

F igure 4.—. From: Point Mutations in the Stem Region and the Fourth AAA Domain of Cytoplasmic Dynein Heavy Chain Partially Suppress the Phenotype of NUDF/LIS1 Loss in Aspergillus nidulans.

Images of GFP-NUDA in different strain backgrounds showing that the nudAR3086C mutation affects dynein localization. Dynein proteins form comet-like structures representing their microtubule-plus-end localization in wild-type cells (wild type). These comet-like structures are more prominent in the ΔnudF mutant (ΔnudF). In the presence of the nudAR3086C mutation, dynein proteins are localized along microtubule-like structures with or without the ΔnudF mutation (ΔnudF/nudAR3086C or nudAR3086C). A merged image is presented to show colocalization of the mutant GFP-NUDA with the CFP microtubule, which is pseudocolored red (nudAR3086C/MT). Bar, ∼5 μm.

Lei Zhuang, et al. Genetics. 2007 March;175(3):1185-1196.
4.
F <span style="font-variant: small-caps" class="small-caps">igure</span>  2.—

F igure 2.—. From: Point Mutations in the Stem Region and the Fourth AAA Domain of Cytoplasmic Dynein Heavy Chain Partially Suppress the Phenotype of NUDF/LIS1 Loss in Aspergillus nidulans.

Growth phenotypes of various nud mutant and nudF suppressor strains at 32° (A) or at 42° (B–D) for 3 days. (A and B) Both the nudAR3086C and the nudAL1098F mutations suppressed the colony phenotype of the nudF deletion mutant (ΔnudF) at 32° (A) and at 42° (B) Note that the nudAR3086C mutation acted as a strong suppressor at both temperatures. (C) The nudAR3086C mutant had a mild growth defect, but the nudAL1098F did not exhibit any obvious colony phenotype. Note that the nudAR3086C mutation also acted as a strong suppressor for the nudF6 mutant. (D) Neither the nudAR3086C mutation nor the nudAL1098F mutation suppressed the growth phenotypes of the ts nudI416 (in dynein intermediate chain) and nudK317 (in Arp1 of the dynactin complex) mutants. Note that the nudAR3086C mutation even deteriorated the slow-growth phenotype of the nudK317 mutant. In A, B, and D, GFP is linked to nudA in all strains except the ΔnudA strain.

Lei Zhuang, et al. Genetics. 2007 March;175(3):1185-1196.
5.
F <span style="font-variant: small-caps" class="small-caps">igure</span>  5.—

F igure 5.—. From: Point Mutations in the Stem Region and the Fourth AAA Domain of Cytoplasmic Dynein Heavy Chain Partially Suppress the Phenotype of NUDF/LIS1 Loss in Aspergillus nidulans.

The ATPase activity of dynein is decreased in the strain carrying the nudAR3086C mutation. (A) A silver-stained gel showing the S-peptide-eluted fractions from strains carrying the S-tagged IC (lane 4 is a negative control without S-IC). Lane 1, nudAL1098C; lane 2, wild type; lane 3, nudAR3086C. All these four strains contain GFP linked with nudA. (B) Relative values of dynein's ATPase activity of the suppressor mutant strains expressed as percentage of the wild-type value. Means and standard errors were calculated from multiple experiments (n = 5 for wild type and nudAR3086C; n = 4 for nudAL1098F). In every experiment starting from protein purification, two or three samples from the same eluted fraction were subjected to ATPase assays and a mean value was obtained and used for further calculations. (C) Estimated values of dynein's ATPase activity. These values were from a single experiment with duplicated ATPase assays. The asterisk indicates that these values are not absolute since the concentration of the dynein heavy chain was estimated from the intensity of a series of BSA samples with different concentrations loaded on the same silver-stained gel. (D). Western blots showing that the levels of dynein HC and NUDF pulled down by the S-IC are not significantly affected by the suppressor mutations.

Lei Zhuang, et al. Genetics. 2007 March;175(3):1185-1196.

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