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. 2002 Feb 1;16(3):377–387. doi: 10.1101/gad.953002

Figure 3.

Figure 3

Mapping and cloning of the slpr locus reveal that slpr encodes a mixed lineage kinase. (A) Meiotic recombination mapping with recessive markers placed slpr to the right of singed (sn) in the 7DE region of the X chromosome. slpr complements the deletions, Df(1)C128 and Df(1)HA11 (+), but fails to complement Df(1)hl-a and Df(1)GE202 (−). These complementation data further refine the position of the slpr gene to the region between 7D13 and 7E2 excluding 7D18–7D20, which is deleted by Df(1)HA11 but complemented by slpr. Next, recombination mapping with P-element transposon insertions in the region (*) define slpr position within 7D13–7D18 (black box). A total of 2/3319 recombinant males were obtained between slpr and EP1167, whereas 0/2315 recombinants were obtained between slpr and EP1143. These results suggest that slpr is closer to the EP1143 marker, and thus likely to be found to the left of Df(1)HA11. Several genes in the region (Tbh, fs(1)h, mys, smox, Traf2, nAcRe, and Trf2) were useful landmarks to correlate the genetic and physical map with the genome sequence. (B) Among predicted and known genes in the defined region, we found a candidate gene encoding a protein kinase, CG2272. A genomic fragment encompassing CG2272 and closely neighboring CG15339, derived from the P1 clone DS08402, was used for slpr mutant rescue experiments. (C) Presence of the genomic transgene is sufficient to rescue the embryonic dorsal open phenotype of slpr. To identify the slpr mutant chromosome among the experimental population of embryonic cuticles, the slpr chromosome was marked with another recessive cuticle marker, gt.Alone, gt/Y embryos show selective loss of ventral denticles; gt,slpr/Y recombinant embryos display both ventral defects and a large dorsal hole. In the presence of the genomic transgene, P{slpr}, the dorsal open phenotype is rescued, whereas the gt phenotype is unaffected. (D) An EST clone, GH26507, derived from the CG2272 locus is 5129 nucleotides, containing 8 exons (black boxes) and 7 introns (lines) relative to genomic DNA. Genbank accession no. AY045717. The cDNA encodes an ORF of 1148 amino acids (see comment in Materials and Methods) with homology to mixed lineage kinases, which display an N-terminal SH3 domain, a catalytic kinase domain, leucine zipper motifs (LZ), and a Cdc42/Rac-interacting binding motif (C). slpr alleles, 921 and 3P5, each have a mutation within the kinase domain as indicated.