Differential use of protein 4.1 translation initiation sites during erythropoiesis: implications for a mutation-induced stage-specific deficiency of protein 4.1 during erythroid development

J A Chasis; L Coulombel; S McGee; G Lee; G Tchernia; J Conboy; N Mohandas

doi:10.1182/blood.V87.12.5324.bloodjournal87125324

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Differential use of protein 4.1 translation initiation sites during erythropoiesis: implications for a mutation-induced stage-specific deficiency of protein 4.1 during erythroid development

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Differential use of protein 4.1 translation initiation sites during erythropoiesis: implications for a mutation-induced stage-specific deficiency of protein 4.1 during erythroid development

J A Chasis, L Coulombel, S McGee, G Lee, G Tchernia, J Conboy and N Mohandas

Blood, Vol.87(12), pp.5324-5331

06/15/1996

DOI: 10.1182/blood.V87.12.5324.bloodjournal87125324

PMID: 8652848

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Abstract

Expression of multiple protein 4.1 isoforms in erythroid progenitors and in a variety of nonerythroid tissues results from alternative pre- mRNA splicing. In 4.1 pre-mRNA, several translation initiation sites are present; synthesis of isoforms larger than 80 kD occurs when an upstream 5′ AUG is spliced in, whereas the 80-kD mature erythroid isoform is produced when the upstream AUG is spliced out and translation is initiated at the downstream AUG. During erythropoiesis, this splicing switch is developmentally regulated. We studied this developmental switch in hereditary elliptocytosis 4.1Alg, in which a DNA rearrangement involving the exon containing the downstream AUG results in loss of coding capacity for the 80-kD 4.1, leading to mature red blood cells deficient in 4.1 with decreased membrane mechanical stability. Analysis of erythroblast RNA by reverse transcriptase- polymerase chain reaction showed that, although it retained the upstream AUG, its coding region was approximately 2.2 kb, compared with approximately 2.5 kb of normal 4.1 mRNA, because of the deletion of exons, including the one that codes for the downstream AUG. Immunofluorescent microscopy and Western blot analysis documented protein 4.1 expression in HE 4.1Alg erythroblasts. These studies emphasize the crucial role of differentiation-regulated RNA splicing because, within the same erythroid tissue, the HE 4.1Alg phenotype did not appear until after the differentiation-associated splicing event.

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Title: Subtitle: Differential use of protein 4.1 translation initiation sites during erythropoiesis: implications for a mutation-induced stage-specific deficiency of protein 4.1 during erythroid development
Creators: J A Chasis - Life Sciences Division, Lawrence Berkeley Laboratory, University of California, Berkeley, CA 94720, USA
L Coulombel - Life Sciences Division, Lawrence Berkeley Laboratory, University of California, Berkeley, CA 94720, USA
S McGee - Life Sciences Division, Lawrence Berkeley Laboratory, University of California, Berkeley, CA 94720, USA
G Lee - Life Sciences Division, Lawrence Berkeley Laboratory, University of California, Berkeley, CA 94720, USA
G Tchernia - Life Sciences Division, Lawrence Berkeley Laboratory, University of California, Berkeley, CA 94720, USA
J Conboy - Life Sciences Division, Lawrence Berkeley Laboratory, University of California, Berkeley, CA 94720, USA
N Mohandas - Life Sciences Division, Lawrence Berkeley Laboratory, University of California, Berkeley, CA 94720, USA
Resource Type: Journal article
Publication Details: Blood, Vol.87(12), pp.5324-5331
DOI: 10.1182/blood.V87.12.5324.bloodjournal87125324
PMID: 8652848
ISSN: 0006-4971
eISSN: 1528-0020
Language: English
Date published: 06/15/1996
Academic Unit: Iowa Neuroscience Institute; Immunology; Internal Medicine
Record Identifier: 9984070294802771

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