Fig. 4. tlp-1 cloning and the tlp-1 locus molecular analysis. (A) tlp-1 was mapped to LG IV between unc-26 and egl-23. Tested genomic clones are shown in relative positions below the genetic map. Red lines represent genomic clones that rescued the tlp-1 T cell defect, in which the number of transgenic lines that rescued the T cell defect is indicated (>70% of the transgenic animals were rescued). (B) Sequence analysis of genomic DNA and 1664 bp composite cDNA revealed the intron/exon structure shown. Boxes indicate exons: specifically, black closed regions indicate the open reading frame; red closed portions indicate the 3'-UTR. Position of the SL1 (trans-spliced leader sequence) and the poly(A) tail are shown. The positions of the base changes in the mh17, bx85 are indicated. (C) Schematic of the TLP-1 protein. Positions of the conserved SPLALLA, S/T-rich, Q-rich and zinc-finger domains are indicated. (D) Phylogenetic relationships of Sp-type proteins (larger font) and some other C2H2 zinc finger proteins (smaller font) used to root the Sp protein phylogeny. The names of the 45 protein sequences begin with a three-letter abbreviation of the organism binomen: Rno, Rattus norvegicus; Mmu, Mus musculus; Hsa, Homo sapiens; Dme, Drosophila melanogaster; Dre, Danio rerio; Cel, Caenorhabditis elegans; Sce, Saccharomyces cerevisiae. Only the least ambiguously aligned residues were used in the analyses. Relationships shown are only those supported with some degree of confidence (multifurcations represent ambiguous branching order, not simultaneous divergence). Numbers (1-24) identify branches with such support in at least one of several different kinds of phylogenetic analyses (see Materials and Methods). All proteins deriving from branch 12 uniquely share the SPLALLA motif, which we therefore propose is a shared-derived feature of these proteins. Branch lengths represent the minimum number of amino acid-altering nucleotide substitutions mapped onto the phylogeny by parsimony.