Amino acid replacement: P162L.
Nucleotide substitution: C834T.
C19184991T
C834T
P162L | gammaTub37C-PA; P162L | gammaTub37C-PB
P162L
Homozygous females lay eggs which appear externally wild type, but fail to hatch. The embryos derived from the homozygous females arrest with one or a few spindle-like structures, which are often large, wide and have chromosomes or chromosome fragments distributed across the spindle structure, rather than being aligned on the metaphase plate.
The chiasmate chromosomes often fail to form a single mass at the spindle midzone in homozygous prometaphase I oocytes, and are instead stretched across the length of the spindle. The mutant chromosomes sometimes appear rounded and misshapen. In some cases, the chromosomes fail to align in any direction and display both abnormal invaginations and projections away from the main chromosome mass. Defects in the DNA threads that connect achiasmate chromosomes are also seen in the mutant oocytes. 70% of the mutant oocytes either lack a recognisable spindle or have an abnormal spindle; monopolar spindles are seen in 22% of cases, barrel-like spindles are seen in 23% of cases and in 15% of cases, the microtubules in the oocyte show no clear directionality and project in all directions.
Dynamic changes in spindle and chromosome configurations are seen in live imaging of mutant oocytes, in contrast to wild-type oocytes where the spindle and chromosomes remain stable for over an hour of observation.
Chromosomes are successfully congressed to the metaphase plate to form lemon-shaped structures in 62% of mutant metaphase I oocytes. However, the chromosomes fail to properly orient in preparation for segregation at anaphase I. 70% of the mutant oocytes have spindles resembling those seen in wild type. The spindle is reduced to a large microtubule bundle projecting from each end of the chromosome mass in 21% of cases.
The nucleus is mislocalised in 22% of mutant oocytes during stages 11 and 12 and is mislocalised in 40% of mutant oocytes during stages 13 and 14.