Nucleotide substitution: A1450T.
The predicted protein would lack its terminal 233 amino acids, including the potential nuclear localization signals and the polar C-terminus.
Amino acid replacement: K464term.
No defect detectable by Southern blotting in the 8.5kb SalI fragment corresponding to that included in P{fs-1}.
A2725790T
A1450T
K464term | fs(1)Ya-PA
K464term
Meiotic chromosome segregation occurs properly and is completed in the absence of fs(1)Ya function in unfertilised eggs.
Two abnormal nuclear distributions are seen in unfertilised eggs from fs(1)Ya2 females. 34% of eggs contain two diploid nuclei, which is never seen in wildtype eggs. 14% of eggs have one diploid nuclei and two haploid nuclei. Of this latter case, two subtypes are observed. In one type, meiosis II has been completed but the central two nuclei, products of a separate meiosis II spindle, subsequently associate. In the other type, meiosis completes normally but subsequently two resulting haploid meiotic products associate or fuse.
There is no differences in the distribution of meiotic stages observed of nuclei upon in vitro activation of unfertilised wildtype vs fs(1)Ya2-deficient eggs.
The majority of fs(1)Ya2 mutant embryos show disrupted nuclear arrangements. In 52% of embryos the male pronucleus does not associate with a maternal nucleus. In 13% of embryos, the male pronucleus does associate with a maternally-derived diploid nucleus in a manner that is never seen in wildtype. 17% of fs(1)Ya2 embryos are in various stages of deterioration with fragmented or dramatically decondensed nuclei and can not be analysed further. 18% of fs(1)Ya2 embryos resemble controls, having a gonomeric nucleus with a haploid maternal content and haploid paternal content, and three haploid maternally derived nuclei.
Roughly 25% of unfertilised fs(1)Ya2 eggs show cell-cycle asynchrony of nuclear chromatin, which is never seen in wildtype. This defect is also observed in a smaller proportion of mutant embryos.
DNA replication can occur in fs(1)Ya2 embryos and eggs.
Mutant embryos arrest either with multiple abnormal nuclei or at the pronuclear stage, with fewer than four abnormally condensed nuclei, presumably derived from the sperm and female meiotic products. In unfertilized eggs, two rather than four nuclei that would go on to form the polar bodies form, and the two are of unequal size and may show different degrees of chromatin condensation. Post meiotic coordination between chromatin condensation and nuclear fusion is abnormal. Fusion into one nucleus eventually occurs but the chromatin looks abnormally condensed.
Following pronuclear migration, 77% +/- 12% of eggs from homozygous fs(1)Ya2 females are arrested with their two pronuclei closely apposed. The DNA in these embryos appears uncondensed, suggesting that the pronuclei are blocked after migration, but before entering the first embryonic mitosis. When these embryos are incubated for longer, their pronuclear DNA disperses and is eventually degraded. The remaining 23% of embryos (escapers) can pass through this initial block and undergo further mitoses, but these mitoses are highly aberrant. Most of these escapers arrest as syncytial embryos, with the remaining 1% arresting as gastrulae.
fs(1)Ya2 is rescued by fs(1)Ya+t8.1
fs(1)Ya2 is rescued by fs(1)Ya+t7.2
fs(1)Ya2 is rescued by fs(1)Ya+t7.7
fs(1)Ya2 is rescued by fs(1)YaFAPKK.Hsp26
fs(1)Ya2 is rescued by fs(1)YaSY160.Hsp26
fs(1)Ya2 is rescued by fs(1)YaHsp26.PL
fs(1)Ya2 is rescued by fs(1)YaFEPKK.Hsp26
fs(1)Ya2 is rescued by fs(1)YaN1.Hsp26.Tag:HA
fs(1)Ya2 is rescued by fs(1)YaΔQ.Hsp26
fs(1)Ya2 is rescued by fs(1)YaΔFSPKK.Hsp26
fs(1)Ya2 is rescued by fs(1)YaHsp26.PL
fs(1)Ya2 is rescued by fs(1)Ya+t8.1
fs(1)Ya2 is rescued by fs(1)Ya+tXSR
fs(1)Ya2 is rescued by fs(1)Ya+t7.2
fs(1)Ya2 is rescued by fs(1)Ya+t8.1
fs(1)Ya2 is partially rescued by fs(1)YaN3.Hsp26.Tag:HA
fs(1)Ya2 is partially rescued by fs(1)YaΔQ.ΔS:T.Hsp26
fs(1)Ya2 is partially rescued by fs(1)YaC3.Hsp26.Tag:HA
fs(1)Ya2 is partially rescued by fs(1)YaΔITPIR.ΔFSPKK.Hsp26
fs(1)Ya2 is partially rescued by fs(1)YaIAPIR.Hsp26
fs(1)Ya2 is partially rescued by fs(1)YaΔS:T.Hsp26
fs(1)Ya2 is partially rescued by fs(1)YaΔITPIR.Hsp26
fs(1)Ya2 is not rescued by fs(1)Yat3.9
fs(1)Ya2 is not rescued by fs(1)Yat6.4
fs(1)Ya2 is not rescued by fs(1)YaC2.Hsp26
fs(1)Ya2 is not rescued by fs(1)YaC2.SY160.Hsp26
fs(1)Ya2 is not rescued by fs(1)YaC1.C2.Hsp26
fs(1)Ya2 is not rescued by fs(1)YaC2.FAPKK.Hsp26
fs(1)Ya2 is not rescued by fs(1)YaC1.C2.FAPKK.Hsp26
fs(1)Ya2 is not rescued by fs(1)YaIAPIR.FAPKK.Hsp26
fs(1)Ya2 is not rescued by fs(1)YaC1.Hsp26
fs(1)Ya2 is fully rescued by fs(1)YaHsp26.PL, fs(1)YaN1.Hsp26.T:Ivir\HA1, fs(1)YaSY160.Hsp26, fs(1)YaΔQ.Hsp26, fs(1)YaΔFSPKK.Hsp26, fs(1)YaFAPKK.Hsp26 and fs(1)YaFEPKK.Hsp26 under both heat shock and non-heat shock conditions. fs(1)Ya2 is not rescued by fs(1)YaC1.Hsp26, fs(1)YaC2.Hsp26, fs(1)YaIAPIR.Hsp26, fs(1)YaC1.C2.Hsp26, fs(1)YaC2.SY160.Hsp26, fs(1)YaC2.FAPKK.Hsp26, fs(1)YaΔITPIR.ΔFSPKK.Hsp26, fs(1)YaIAPIR.FAPKK.Hsp26, fs(1)YaC1.C2.FAPKK.Hsp26 or fs(1)Ya4.28.1.1 under either heat shock and non-heat shock conditions. fs(1)Ya2 is not rescued under non-heat shock conditions, and very weakly rescued under heat shock conditions by fs(1)YaN3.Hsp26.T:Ivir\HA1, fs(1)YaΔS:T.Hsp26 or fs(1)YaC3.Hsp26.T:Ivir\HA1. fs(1)Ya2 is slightly rescued by fs(1)YaΔITPIR.Hsp26 and fs(1)YaΔQ.ΔS:T.Hsp26 under both heat shock and non-heat shock conditions.
Mutant phenotype can be rescued by the fs(1)YaHsp26.PL construct, females are fertile.
Young.
fs(1)Ya2 is an amorph, as the phenotype of homozygous fs(1)Ya2 females is the same as hemizygous fs(1)Ya2/Df(1)w-rJ1 flies, both in terms of their fecundity and the embryonic phenotype of their progeny. However fs(1)Ya2 makes normal amounts of fs(1)Ya2 RNA and protein of expected sizes.