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An Erythroid-Specific Transcript Generates the Soluble Form of
NADH-Cytochrome b5 Reductase in Humans
Alessandra Bulbarelli,
Alessandra Valentini,
Marcella DeSilvestris,
M. Domenica Cappellini, and
Nica Borgese
From Consiglio Nazionale delle Ricerche Center for Cellular and
Molecular Pharmacology, Department of Pharmacology, University of
Milan, Milan; Faculty of Pharmacy, University of Catanzaro, Catanzaro;
Bayer Research, Biological and Technological Research Department,
Milan; and Centro Anemie Congenite-Ospedale Maggiore Policlinico IRCCS,
University of Milan, Milan, Italy.
Two forms of NADH-cytochrome b5 reductase (b5R), an
erythrocyte-restricted soluble form, active in methemoglobin reduction, and a ubiquitous membrane-associated form involved in lipid metabolism, are produced from one gene. In the rat, the two forms are generated from alternative transcripts differing in the first exon, however, biogenesis of human b5R was less understood. Recently, two different transcripts (M and S), differing in the first exon were also described in humans. Here, we have investigated the
tissue-specificity and the role of the S-transcript in the generation
of soluble b5R. By RNase protection assays designed to simultaneously
detect alternative b5R transcripts in the same sample, the S transcript
was undetectable in nonerythroid and in erythroleukemic K562 cells
induced to differentiate, but was present in terminal erythroblast
cultures, and represented a major b5R transcript in reticulocytes.
Analysis of the translation products of the M- and S-transcripts in
HeLa cells transfected with the corresponding cDNAs demonstrated that
the S-transcript generates soluble b5R, presumably from an internal
initiation codon. Our results indicate that the
S-transcript is expressed at late stages of erythroid maturation to
generate soluble b5R.
Blood, Vol. 92 No. 1 (July 1), 1998:
pp. 310-319
© 1998 by The American Society of Hematology.
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