Volume 8, Issue 2, Pages 375-381 (August 2001) A Human Homolog of Yeast Pre-mRNA Splicing Gene, PRP31, Underlies Autosomal Dominant Retinitis Pigmentosa on Chromosome 19q13.4 (RP11)  Eranga N. Vithana, Leen Abu-Safieh, Maxine J. Allen, Alisoun Carey, Myrto Papaioannou, Christina Chakarova, Mai Al-Maghtheh, Neil D. Ebenezer, Catherine Willis, Anthony T. Moore, Alan C. Bird, David M. Hunt, Shomi S. Bhattacharya  Molecular Cell  Volume 8, Issue 2, Pages 375-381 (August 2001) DOI: 10.1016/S1097-2765(01)00305-7

Figure 1 Physical Map of the RP11 Interval and the Genomic Organization of PRPF31 STSs, ESTs, and genes mapping within the refined interval D19S927 and D19S781.2 were used to construct the physical map based on PAC and BAC clones. Only a few of the clones of the contig are presented; ones being sequenced by LLNL are numbered in red. The genes excluded from RP causation, including NIX-predicted genes (WI-17997, CACNG5, L-CACNG, KIAA0691, FB1, BB1, and A006I07), are shown in red. An enlarged view of the PRPF31 gene is shown. It consists of 14 coding exons between 61 and 185 bp in length, with the initiation and stop codons (ATG and TAG) present in exons 2 and 14, respectively. The first intron is within the 5′ UTR. All exons follow the GT/AG rule (Breathnach and Chambon, 1981). Sizes of exons and introns are drawn to scale Molecular Cell 2001 8, 375-381DOI: (10.1016/S1097-2765(01)00305-7)

Figure 2 Expression of PRPF31 cDNA expression profile for exons 13 and 14 of PRPF31 showing amplification of the 350-bp product from all tissues including retina. A ubiquitously expressed gene, PGM1 (Whitehouse et al., 1992), was used as a control; its 400-bp fragment was also observed in all tissues Molecular Cell 2001 8, 375-381DOI: (10.1016/S1097-2765(01)00305-7)

Figure 3 Cosegregation Analysis of the Mutations (A) Segregation of the 43-bp deletion mutation (IVS6-3 to -45 del) in pedigree RP677 as shown by agarose gel electrophoresis. Lane assignment (1–10) corresponds to samples of the respective individuals in the pedigree. Individual 7 in the pedigree is an asymptomatic obligate carrier and demonstrates the incomplete penetrance in this pedigree. All affected individuals exhibit the deleted allele. (B) Segregation of the 11-bp deletion mutation (1115–1125 del) in pedigree AD5. Only a branch of AD5 pedigree is shown along with the haplotypes for the RP11 markers; the asterisk denotes individual II-2 in the original pedigree (Al-Maghtheh et al., 1994). Lane assignment (1–22) corresponds to samples of the respective individuals in the pedigree. Individuals 12 and 15 are asymptomatic obligate carriers, while individuals 3, 9, 14, and 22 denote asymptomatic disease haplotype carriers. All affected haplotype carriers exhibit the deleted allele Molecular Cell 2001 8, 375-381DOI: (10.1016/S1097-2765(01)00305-7)

Figure 4 Protein Alignment of Human PRPF31 (DKFZP566JI53 Protein, Residues 95 to 383), PRP31-like Proteins from Other Species, and Nop, a Putative snoRNA Binding Domain PRPF31 protein shows highest homologies over its full-length to the CG6876 gene product of D. melanogaster (60% over the entire protein), A. thaliana hypothetical protein T13D8.1 (45% from residues 2 to 490), prp31+ from S. pombe (38% from residues 1 to 477), and Prp31p from S. cerevisiae (31% from residues 97 to 426). Amino acids, which exhibit complete identity across species, are boxed, whereas amino acids identical to those of the Nop snoRNA domain are shown in red. The region between residues 351 and 364 of PRPF31 (highlighted in blue) with the consensus sequence 351(R/K)(R/K)(R/K) R XX(R/K)(R/K)XRKXKE364 harbors three putative nuclear localization signals (NLS) in agreement with the proposed consensus sequence, K(K/R)X(K/R), for monopartite NLSs (Chelsky et al., 1989). Red circles indicate the most likely signal sequence of the three according to PSORT, a program that identifies subcellular localization signals. Blue diamonds indicate the amino acid residues 194 and 216 mutated in sporadic case 42 (SP42) and AD29, respectively. In individual SP14, the underlined residues ELERLEEACDM are duplicated due to mutation (580–581 dup 33 bp). In individual SP117, the insertion mutation (769–770 ins A) adds 20 novel amino acids after codon 256 followed by a premature stop in codon 278; the novel residues of this protein are shown in mauve. In AD5, the deletion mutation (1115–1125 del) adds 98 novel amino acids after codon 371 followed by a premature stop in codon 470; the first 13 novel residues of the aberrant protein are shown in mauve Molecular Cell 2001 8, 375-381DOI: (10.1016/S1097-2765(01)00305-7)