EMS-generated point mutation in Chro that introduces a premature stop codon resulting in a truncated 71 amino acid protein. The truncated N-terminal fragment does not contain the chromodomain.
Nucleotide substitution: G402A
In contrast to wild-type, in Chro71/Chro612 mutant larvae at 96h after larval hatching (ALH) 26% of mira-positive central brain neural stem cells (NSCs) retain their cellular processes and number of actively dividing dpn-positive NSCs is much reduced compared to controls (the cells however do not undergo premature differentiation as judged by expression of various markers). In wild-type larvae all central brain neuroblasts reactivate (lose the cellular process and are mitotically active) by 48h ALH, whereas in Chro71/Chro612 mutants large proportion of the neuroblasts are still quiescent. Neuroblast reactivation defects are also observed in the thoracic ventral nerve cord NSCs Chro71/Chro17a as well as Chro71/Chro8c mutants all display similar defects in the reactivation of central brain neuroblasts.
90% of Chro71/Chro612 mutant larval neuroblasts have severe spindle and chromosome segregation defects. The microtubule spindles are incomplete, unfocussed, and/or without clear spindle poles. At anaphase, chromosomes are lagging and scattered.
The mitotic index is significantly reduced in Chro71/Chro612 mutants, and an associated small brain and imaginal disc phenotype is observed.
Chro71/Chro612 has lethal | third instar larval stage phenotype, enhanceable by JIL-1z2
Chro71/Chro612 has decreased occurrence of cell division | third instar larval stage phenotype, suppressible | partially by Scer\GAL4insc-Mz1407/grhUAS.cUa
Chro71/Chro612 has decreased occurrence of cell division | third instar larval stage phenotype, suppressible | partially by pros[+]/pros17
Chro71/Chro612, pros[+]/pros17 has decreased occurrence of cell division | third instar larval stage phenotype, suppressible | partially by Scer\GAL4insc-Mz1407/grhUAS.cUa
Chro71/Chro612, Scer\GAL4insc-Mz1407, grhUAS.cUa has decreased occurrence of cell division | third instar larval stage phenotype, suppressible | partially by pros[+]/pros17
Chro71/Chro612 has neuroblast | third instar larval stage phenotype, suppressible | partially by Scer\GAL4insc-Mz1407/grhUAS.cUa
Chro71/Chro612 has neuroblast | third instar larval stage phenotype, suppressible | partially by pros[+]/pros17
Chro71/Chro17a has neuroblast | third instar larval stage phenotype, suppressible | partially by pros[+]/pros17
Chro71/Chro8c has neuroblast | third instar larval stage phenotype, suppressible | partially by pros[+]/pros17
Chro71/Chro612, pros[+]/pros17 has neuroblast | third instar larval stage phenotype, suppressible | partially by Scer\GAL4insc-Mz1407/grhUAS.cUa
Chro71/Chro612, Scer\GAL4insc-Mz1407, grhUAS.cUa has neuroblast | third instar larval stage phenotype, suppressible | partially by pros[+]/pros17
The central brain neuroblast reactivation defects (manifested by the retention of cellular extensions and low mitotic activity even at late larval stages) characteristic for Chro71/Chro612 mutants are alleviated by expression of grhScer\UAS.cUa under the control of Scer\GAL4insc-Mz1407 in the mutant background or by combination with a single copy of pros17, the phenotype can be further improved when both grhScer\UAS.cUa expression as well as the pros17 heterozygosity are combined. The neuroblast cellular extension phenotype of Chro71/Chro17a as well as Chro71/Chro8c larvae is also significantly suppressed by pros17 heterozygosity.
Chro71/Chro612 is rescued by Chro+tCH322-159M1
Chro71/Chro612 is partially rescued by Scer\GAL4unspecified/ChroNTD.UASp.GFP.Tag:NLS(SV40-largeT)
Chro71/Chro612 is partially rescued by Scer\GAL4elav-C155/ChroGFP
Chro71/Chro612 is not rescued by Scer\GAL4unspecified/ChroNTD-ΔChD.UAS.Tag:NLS(Unk),Tag:V5
Chro71/Chro612 is not rescued by Scer\GAL4unspecified/ChroChD.UAS.Tag:NLS(SV40-largeT),Tag:V5,GFP
Chro71/Chro612 is not rescued by Scer\GAL4unspecified/ChroCTD.UASp.GFP
The brain neuroblast reactivation defects observed in either Chro71/Chro612 or Chro612/Df(3L)ED231 mutant larvae at 96h after larval hatching can be rescued by combination with Chro+tCH322-159M1.
Expression of ChroScer\UAS.P\T.FL.T:Ivir\HA1,T:Zzzz\FLAG,T:Avic\GFP in third larval instar salivary glands rescues all aspects of polytene chromosome morphology in Chro71/Chro612 animals.
Expression of ChroNTD.Scer\UAS.P\T.T:Avic\GFP.T:SV40\nls2 in third larval instar salivary glands substantially rescues polytene chromosome morphology in Chro71/Chro612 animals, although some regions of the chromosome arms remain coiled.
Expression of either ChroNTD-ΔChD.Scer\UAS.T:Uuuu\nls6,T:SV5\V5 or ChroChD.Scer\UAS.T:SV40\nls2,T:SV5\V5,T:Avic\GFP in third larval instar salivary glands results in no or very little improvement of polytene chromosome morphology in Chro71/Chro612 animals.
Expression of ChroCTD.Scer\UAS.P\T.T:Avic\GFP in third larval instar salivary glands does not rescue polytene chromosome morphology in Chro71/Chro612 animals.
Expression of ChroScer\UAS.P\T.T:Avic\GFP with Scer\GAL4elav-C155 partially rescues the smaller brain size, and coiled and condensed chromosome arm morphology of the polytene chromosomes from Chro71/Chro612 mutant larvae.