{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,2]],"date-time":"2026-05-02T04:26:31Z","timestamp":1777695991737,"version":"3.51.4"},"reference-count":59,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2004,3,26]],"date-time":"2004-03-26T00:00:00Z","timestamp":1080259200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Br J Haematol"],"published-print":{"date-parts":[[2004,4]]},"abstract":"<jats:title>Summary<\/jats:title><jats:p>Two single\u2010nucleotide substitutions in <jats:italic>PKLR<\/jats:italic> constituted the molecular basis underlying pyruvate kinase (PK) deficiency in a patient with severe haemolytic anaemia. One novel mutation, IVS5+1G&gt;A, abolished the intron 5 donor splice site. The other mutation, c.1436G&gt;A, altered the intron 10 donor splice site consensus sequence and, moreover, encoded an R479H substitution. We studied the effects on <jats:italic>PKLR<\/jats:italic> pre\u2010mRNA processing, using <jats:italic>ex vivo<\/jats:italic>\u2010produced nucleated erythroid cells from the patient. Abolition of the intron 5 splice site initiated two events in the majority of transcripts: skipping of exon 5 or, surprisingly, simultaneous skipping of exon 5 and 6 (\u03945,6). Subcellular localization of transcripts suggested that no functional protein was produced by the IVS5+1A allele. The unusual \u03945,6 transcript suggests that efficient inclusion of exon 6 in wild\u2010type <jats:italic>PKLR<\/jats:italic> mRNA depends on the presence of splice\u2010enhancing elements in exon 5. The c.1436G&gt;A mutation caused skipping of exon 10 but was mainly associated with a severe reduction in transcripts although these were, in general, normally processed. Accordingly, low amounts of PK were detected in nucleated erythroid cells of the patient, thus correlating with the patient's PK\u2010deficient phenotype. Finally, several low\u2010abundant transcripts were detected that represent the first examples of \u2018leaky\u2010splicing\u2019 in <jats:italic>PKLR<\/jats:italic>.<\/jats:p>","DOI":"10.1111\/j.1365-2141.2004.04895.x","type":"journal-article","created":{"date-parts":[[2004,6,18]],"date-time":"2004-06-18T13:36:21Z","timestamp":1087565781000},"page":"253-263","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["<i>Ex vivo<\/i> analysis of aberrant splicing induced by two donor site mutations in <i>PKLR<\/i> of a patient with severe pyruvate kinase deficiency"],"prefix":"10.1111","volume":"125","author":[{"given":"Richard","family":"van Wijk","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Annet C. W.","family":"van Wesel","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Adri A. M.","family":"Thomas","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gert","family":"Rijksen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wouter W.","family":"van Solinge","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2004,3,26]]},"reference":[{"key":"e_1_2_20_2_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V98.13.3685"},{"key":"e_1_2_20_3_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.90.9.4324"},{"key":"e_1_2_20_4_1","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1016\/S0308-2261(21)00623-8","article-title":"Metabolism in haemolytic states","volume":"4","author":"Ben\u00f6hr H.C.","year":"1975","journal-title":"Clinical Haematology"},{"key":"e_1_2_20_5_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.270.6.2411"},{"key":"e_1_2_20_6_1","volume-title":"Red Cell Metabolism: A Manual of Biochemical Methods","author":"Beutler E.","year":"1984"},{"key":"e_1_2_20_7_1","doi-asserted-by":"publisher","DOI":"10.1006\/bcmd.2000.0276"},{"key":"e_1_2_20_8_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.biochem.72.121801.161720"},{"key":"e_1_2_20_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0968-0004(00)01549-8"},{"key":"e_1_2_20_10_1","first-page":"525","volume-title":"The RNA World","author":"Burge C.B.","year":"1999"},{"key":"e_1_2_20_11_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrg775"},{"key":"e_1_2_20_12_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2141.1998.01094.x"},{"key":"e_1_2_20_13_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V92.2.647.414k13_647_652"},{"key":"e_1_2_20_14_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1073774"},{"key":"e_1_2_20_15_1","doi-asserted-by":"publisher","DOI":"10.1101\/gad.1048803"},{"key":"e_1_2_20_16_1","doi-asserted-by":"publisher","DOI":"10.1093\/hmg\/8.10.1893"},{"key":"e_1_2_20_17_1","first-page":"5b","article-title":"Exonic point mutation 1552 CGT\u2192AGT is not a missense mutation but causes an aberrant splicing of human L\u2010type pyruvate kinase gene","volume":"90","author":"Fujii H.","year":"1997","journal-title":"Blood"},{"key":"e_1_2_20_18_1","doi-asserted-by":"publisher","DOI":"10.1017\/S1355838200000960"},{"key":"e_1_2_20_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0955-0674(00)00212-X"},{"key":"e_1_2_20_20_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.genet.33.1.229"},{"key":"e_1_2_20_21_1","first-page":"4637","volume-title":"The Metabolic and Molecular Bases of Inherited Disease","author":"Hirono A.","year":"2001"},{"key":"e_1_2_20_22_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2141.1985.tb04059.x"},{"key":"e_1_2_20_23_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0076-6879(82)90119-7"},{"key":"e_1_2_20_24_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V84.10.3505.3505"},{"key":"e_1_2_20_25_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V89.11.4213"},{"key":"e_1_2_20_26_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.bi.65.070196.002055"},{"key":"e_1_2_20_27_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF00210743"},{"key":"e_1_2_20_28_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1098-1004(200003)15:3<261::AID-HUMU7>3.0.CO;2-T"},{"key":"e_1_2_20_29_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.97.17.9591"},{"key":"e_1_2_20_30_1","doi-asserted-by":"publisher","DOI":"10.1017\/S1355838202028030"},{"key":"e_1_2_20_31_1","doi-asserted-by":"crossref","first-page":"2817","DOI":"10.1182\/blood.V83.10.2817.2817","article-title":"Mutations in the pyruvate kinase L gene in patients with hereditary hemolytic anemia","volume":"83","author":"Lenzner C.","year":"1994","journal-title":"Blood"},{"key":"e_1_2_20_32_1","doi-asserted-by":"publisher","DOI":"10.1101\/gad.12.13.1998"},{"key":"e_1_2_20_33_1","doi-asserted-by":"publisher","DOI":"10.1038\/83762"},{"key":"e_1_2_20_34_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2141.2000.02283.x"},{"key":"e_1_2_20_35_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.0007-1048.2002.03665.x"},{"key":"e_1_2_20_36_1","doi-asserted-by":"publisher","DOI":"10.1038\/416499a"},{"key":"e_1_2_20_37_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(01)00202-1"},{"key":"e_1_2_20_38_1","doi-asserted-by":"publisher","DOI":"10.1016\/0006-291X(79)90592-8"},{"key":"e_1_2_20_39_1","doi-asserted-by":"publisher","DOI":"10.1128\/MCB.19.3.1853"},{"key":"e_1_2_20_40_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(01)00583-9"},{"key":"e_1_2_20_41_1","doi-asserted-by":"publisher","DOI":"10.1006\/bcmd.2002.0502"},{"key":"e_1_2_20_42_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2141.1979.tb03751.x"},{"key":"e_1_2_20_43_1","doi-asserted-by":"publisher","DOI":"10.1016\/0378-1119(94)90567-3"},{"key":"e_1_2_20_44_1","doi-asserted-by":"crossref","first-page":"1871","DOI":"10.1182\/blood.V77.9.1871.1871","article-title":"Point mutations in the L\u2010type pyruvate kinase gene of two children with hemolytic anemia caused by pyruvate kinase deficiency","volume":"77","author":"Neubauer B.","year":"1991","journal-title":"Blood"},{"key":"e_1_2_20_45_1","doi-asserted-by":"publisher","DOI":"10.1038\/ng1129"},{"key":"e_1_2_20_46_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(02)00617-7"},{"key":"e_1_2_20_47_1","doi-asserted-by":"publisher","DOI":"10.1002\/ajh.2830350309"},{"key":"e_1_2_20_48_1","doi-asserted-by":"crossref","first-page":"2895","DOI":"10.1182\/blood.V96.8.2895","article-title":"Nonsense mutations in the human \u03b2\u2010globin gene lead to unexpected levels of cytoplasmic mRNA accumulation","volume":"96","author":"Rom\u00e3o L.","year":"2000","journal-title":"Blood"},{"key":"e_1_2_20_49_1","doi-asserted-by":"publisher","DOI":"10.1128\/MCB.19.1.261"},{"key":"e_1_2_20_50_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/15.17.7155"},{"key":"e_1_2_20_51_1","doi-asserted-by":"publisher","DOI":"10.1006\/mthe.2002.0805"},{"key":"e_1_2_20_52_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/30.9.2083"},{"key":"e_1_2_20_53_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M202107200"},{"key":"e_1_2_20_54_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2002-07-2321"},{"key":"e_1_2_20_55_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2002-06-1851"},{"key":"e_1_2_20_56_1","doi-asserted-by":"crossref","first-page":"2781","DOI":"10.1101\/gad.943701","article-title":"Curbing the nonsense: the activation and regulation of mRNA surveillance","volume":"15","author":"Wilusz C.J.","year":"2001","journal-title":"Genes and Development"},{"key":"e_1_2_20_57_1","first-page":"487","volume-title":"The RNA World","author":"Yu Y.\u2010T.","year":"1999"},{"key":"e_1_2_20_58_1","doi-asserted-by":"crossref","first-page":"3847","DOI":"10.1182\/blood.V89.10.3847","article-title":"Molecular characterization of PK\u2010LR gene in pyruvate kinase\u2010deficient Italian patients","volume":"89","author":"Zanella A.","year":"1997","journal-title":"Blood"},{"key":"e_1_2_20_59_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2141.2001.02711.x"},{"key":"e_1_2_20_60_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature01031"}],"container-title":["British Journal of Haematology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1111%2Fj.1365-2141.2004.04895.x","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1111%2Fj.1365-2141.2004.04895.x","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/j.1365-2141.2004.04895.x","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,16]],"date-time":"2023-10-16T11:06:44Z","timestamp":1697454404000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/j.1365-2141.2004.04895.x"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2004,3,26]]},"references-count":59,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2004,4]]}},"alternative-id":["10.1111\/j.1365-2141.2004.04895.x"],"URL":"https:\/\/doi.org\/10.1111\/j.1365-2141.2004.04895.x","archive":["Portico"],"relation":{},"ISSN":["0007-1048","1365-2141"],"issn-type":[{"value":"0007-1048","type":"print"},{"value":"1365-2141","type":"electronic"}],"subject":[],"published":{"date-parts":[[2004,3,26]]},"assertion":[{"value":"2004-03-26","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}