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CC-5663 ift74-2 IFT74Î”130 ift81-1 IFT81(5E) mt+

Name: CC-5663 ift74-2 IFT74Î”130 ift81-1 IFT81(5E) mt+
SKU: CC-5663
Availability: InStock

$30.00

From George B. Witman, University of Massachusetts Medical School, August 2020

This is a double mutant made by mating ift74-2 IFT74Î”130 (Brown et al., 2015) to ift81-1 IFT81(5E) (Kubo et al., 2016). The strain expresses both IFT74Î”130 and IFT81(5E) in a background otherwise null for IFT74 and IFT81. IFT74Î”130 is a version of IFT74 lacking aa 1-130 important for the proteinâ€™s interaction with the highly acidic tail (also known as E-hook) of Î²-tubulin. IFT81(5E) is a version of IFT81 in which five highly conserved basic residues (K73, R75, R87, K114, and R115) in the proteinâ€™s calponin-homology domain have been replaced by glutamate to reduce or eliminate the proteinâ€™s binding to tubulin. The strain is predicted to lack nearly all intraflagellar transport of tubulin. It has a strong palmelloid phenotype and forms very short flagella with normal axonemal ultrastructure.

It was created by insertional mutagenesis of the parent strains with a fragment conferring resistance to hygromycin B (aph7â€) to generate ift74-2 and ift81-1, followed by transformation with a DNA fragment encoding IFT74Î”130 or IFT81(5E), respectively, and containing a paromomycin-resistance gene as a selectable marker. The transformed strains were then crossed to generate the double mutant.

Mutant alleles: ift74-2, chromosome_1:4204163-4208957; ift81-1, chromosome_17:3362408-3368081

Brown JM, Cochran DA, Craige B, Kubo T, Witman GB. Assembly of IFT trains at the ciliary base depends on IFT74. Curr Biol. 2015 Jun 15;25(12):1583-93. doi: 10.1016/j.cub.2015.04.060. Epub 2015 Jun 4. PMID: 26051893; PMCID: PMC4482480.

Kubo T, Brown JM, Bellve K, Craige B, Craft JM, Fogarty K, Lechtreck KF, Witman GB. Together, the IFT81 and IFT74 N-termini form the main module for intraflagellar transport of tubulin. J Cell Sci. 2016 May 15;129(10):2106-19. doi: 10.1242/jcs.187120. Epub 2016 Apr 11. PMID: 27068536; PMCID: PMC5506485.

Van De Weghe JC, Harris JA, Kubo T, Witman GB, Lechtreck KF. Diffusion rather than IFT likely provides most of the tubulin required for axonemal assembly. J Cell Sci. 2020 Aug 14:jcs.249805. doi: 10.1242/jcs.249805. Epub ahead of print. PMID: 32801124.

Locus:
IFT74, IFT81
Chromosome:
1,17
Mutation Information:
Predicted Mutation Effect:

CC-5663 ift74-2 IFT74Î”130 ift81-1 IFT81(5E) mt+

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Banner Photo Credit: Chlamydomonas reinhardtii zygotes were stained for immunofluorescence with anti-acetylated tubulin (green), anti-phospholipase D (red), and DAPI (blue) by Karl F. Lechtreck (University of Georgia) and George B. Witman (University of Massachusetts Medical School).

Any opinions, findings, and conclusions or recommendations expressed on this website are those of the author(s) and do not necessarily reflect the views of the National Science Foundation.

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