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J. Biol. Chem., Vol. 275, Issue 36, 28301-28307, September 8, 2000
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From the Department of Microbiology and Immunology, Stanford
University School of Medicine, Stanford, California 94305
Analysis by reverse transcription-polymerase
chain reaction has suggested the existence of at least two La
autoantigen-encoding mRNAs that contain different 5' noncoding
regions (NCRs) linked to the same La coding region (Troster, H.,
Metzger, T. E., Semsei, I., Schwemmle, M., Winterpacht, A.,
Zabel, B., and Bachmann, M. (1994) J. Exp. Med.
180, 2059-2067). La-encoding transcripts La1 and La1' contain
115- and 483-nucleotide 5' NCRs, respectively. To determine whether the
various La transcripts are functional mRNAs, the expression and
polysomal association of natural La1 and La1' RNAs were examined.
Although La1 transcripts were ubiquitously expressed in human tissues,
La1' transcripts were predominantly expressed in peripheral blood
leukocytes, especially in B, T, and natural killer cells. Both La1 and
La1' transcripts associated with polysomes in natural killer cells,
suggesting that these transcripts were functional mRNAs. Upon
activation of B cells with the mitogens phorbol 12-myristate 13-acetate
and ionomycin, the amount of La1' mRNA, but not La1, declined. In
contrast, after chemical activation of T cells, the amount of La 1 mRNA, but not La1', declined. The mechanism by which the La1 and
La1' 5' NCRs initiate translation initiation was tested in cultured
human HeLa cells and in two different in vitro translation
systems. It was found that both 5' NCRs can mediate translation
initiation by internal initiation. These findings indicate that the
constitutive expression of La1 mRNA and the tissue-specific
expression of La1' mRNA can both allow La protein synthesis under
conditions when cap-dependent translation is compromised,
such as inflammation, apoptosis, or certain viral infections.
Distinct mRNAs That Encode La Autoantigen Are Differentially
Expressed and Contain Internal Ribosome Entry Sites*
*
This work was supported by National Institutes of Health
Grant R01 GM55979 (to P. S.) and by American Cancer Society Grant PF-98-16-01-MBC (to M. S. C.).The costs of publication of this article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Dept. of Microbiology
and Immunology, Stanford University School of Medicine, Fairchild
Science Bldg., Stanford, CA 94305. Tel.: 650-498-7076; Fax:
650-498-7147; E-mail: psarnow@leland.stanford.edu.
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