Genotype-proteotype linkage in the Wiskott-Aldrich syndrome.
Wiskott-Aldrich syndrome (WAS) is a platelet/immunodeficiency disease arising from mutations of WAS protein (WASP), a hemopoietic cytoskeletal protein. Clinical symptoms vary widely from mild (X-linked thrombocytopenia) to life threatening. In this study, we examined the molecular effects of individual mutations by quantifying WASP in peripheral lymphocytes of 44 patients and identifying the molecular variant (collectively called proteotype). Nonpredicted proteotypes were found for 14 genotypes. These include WASP-negative lymphocytes found for five missense genotypes and WASP-positive lymphocytes for two nonsense, five frameshift, and two splice site genotypes. Missense mutations in the Ena/VASP homology 1 (EVH1) domain lead to decreased/absent WASP but normal mRNA levels, indicating that proteolysis causes the protein deficit. Because several of the EVH1 missense mutations alter WIP binding sites, the findings suggest that abrogation of WIP binding induces proteolysis. Whereas platelets of most patients were previously shown to lack WASP, WASP-positive platelets were found for two atypical patients, both of whom have mutations outside the EVH1 domain. WASP variants with alternative splicing and intact C-terminal domains were characterized for eight nonsense and frameshift genotypes. One of these, a nonsense genotype in a mild patient, supports expression of WASP lacking half of the proline-rich region. With one notable exception, genotype and proteotype were linked, indicating that a genotype-proteotype registry could be assembled to aid in predicting disease course and planning therapy for newly diagnosed infants. Knowledge of the molecular effect of mutations would aid also in identifying disease-modifying genes.
CBR Institute for Biomedical Research, and Department of Pediatrics, Harvard Medical School, 800 Huntington Avenue, Boston, MA 02115, USA.
Maxim I
MI
Lutskiy
Fred S
FS
Rosen
Eileen
E
Remold-O'Donnell
D000293:Adolescent; D000328:Adult; D017398:Alternative Splicing / Q000235:genetics; D001792:Blood Platelets / Q000378:metabolism / Q000473:pathology; D002648:Child; D002675:Child, Preschool; D018389:Codon, Nonsense; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics; D016368:Frameshift Mutation; D008040:Genetic Linkage / Q000276:immunology; D005838:Genotype; D006801:Humans; D007223:Infant; D008297:Male; D020125:Mutation, Missense; D017434:Protein Structure, Tertiary / Q000235:genetics; D011506:Proteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D022821:RNA Splice Sites / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis / Q000378:metabolism; D014923:Wiskott-Aldrich Syndrome / Q000097:blood / Q000235:genetics* / Q000276:immunology; D051316:Wiskott-Aldrich Syndrome Protein
D018389:Codon, Nonsense; D004268:DNA-Binding Proteins; C092744:ENA-VASP proteins; D011506:Proteins; D022821:RNA Splice Sites; D012333:RNA, Messenger; C495767:WAS protein, human; D051316:Wiskott-Aldrich Syndrome Protein