RNA secondary structure:: An important cis-element in rat calcitonin/CGRP pre-messenger RNA splicing

被引:33
作者
Coleman, TP [1 ]
Roesser, JR [1 ]
机构
[1] Virginia Commonwealth Univ, Dept Biochem, Richmond, VA 23227 USA
关键词
D O I
10.1021/bi9808058
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The calcitonin/CGRP gene pre-mRNA is alternatively spliced in a tissue-specific manner resulting in the formation of calcitonin mRNA in thyroid C-cells and CGRP mRNA in neurons. Computer analysis of the RNA containing the 3' splice acceptor of the calcitonin-specific exon 4 predicts that this region has the potential to form a thermodynamically stable stem-loop. Data from CD spectroscopy and solution phase structure probing with single-strand specific and double-strand specific RNases indicates that RNA in this region is substantially double stranded. In vitro splicing of chimeric human beta-globin/calcitonin transcripts in HeLa nuclear extract was inhibited by base changes predicted to destabilize the stem, while compensatory base changes resulted in splicing at 50% of wild-type levels. Changing the residue opposite the AG dinucleotide adenosine in the stem from G to U, allowing the formation of an A-U basepair, abolished usage of this splice acceptor in vitro. These results indicate that a thermodynamically stable RNA stem-loop forms in vitro at the 3' splice acceptor of exon 4 of the calcitonin/CGRP gene transcript. This RNA secondary structure acts as a novel cis-element involved in proper splice site selection.
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收藏
页码:15941 / 15950
页数:10
相关论文
共 44 条
  • [1] IDENTIFICATION OF A NOVEL HIV-1 TAR RNA BULGE BINDING-PROTEIN
    BAKER, B
    MUCKENTHALER, M
    VIVES, E
    BLANCHARD, A
    BRADDOCK, M
    NACKEN, W
    KINGSMAN, AJ
    KINGSMAN, SM
    [J]. NUCLEIC ACIDS RESEARCH, 1994, 22 (16) : 3365 - 3372
  • [2] PREMESSENGER RNA SECONDARY STRUCTURE AND THE REGULATION OF SPLICING
    BALVAY, L
    LIBRI, D
    FISZMAN, MY
    [J]. BIOESSAYS, 1993, 15 (03) : 165 - 169
  • [3] SOLUTION STRUCTURE OF SELENOCYSTEINE-INSERTING TRANSFER RNA(SEC) FROM ESCHERICHIA-COLI - COMPARISON WITH CANONICAL TRANSFER RNA(SER)
    BARON, C
    WESTHOF, E
    BOCK, A
    GIEGE, R
    [J]. JOURNAL OF MOLECULAR BIOLOGY, 1993, 231 (02) : 274 - 292
  • [4] BLACK DL, 1995, RNA, V1, P763
  • [5] Blanchette M, 1997, RNA, V3, P405
  • [6] CHAN RC, 1995, MOL CELL BIOL, V15, P6377
  • [7] IDENTIFICATION OF EXON SEQUENCES AND AN EXON BINDING-PROTEIN INVOLVED IN ALTERNATIVE RNA SPLICING OF CALCITONIN CGRP
    COTE, GJ
    STOLOW, DT
    PELEG, S
    BERGET, SM
    GAGEL, RF
    [J]. NUCLEIC ACIDS RESEARCH, 1992, 20 (09) : 2361 - 2366
  • [8] CALCITONIN EXON SEQUENCES INFLUENCE ALTERNATIVE RNA PROCESSING
    COTE, GJ
    NGUYEN, IN
    BERGET, SM
    GAGEL, RF
    [J]. MOLECULAR ENDOCRINOLOGY, 1990, 4 (11) : 1744 - 1749
  • [9] NEURON-SPECIFIC ALTERNATIVE RNA PROCESSING IN TRANSGENIC MICE EXPRESSING A METALLOTHIONEIN CALCITONIN FUSION GENE
    CRENSHAW, EB
    RUSSO, AF
    SWANSON, LW
    ROSENFELD, MG
    [J]. CELL, 1987, 49 (03) : 389 - 398
  • [10] DABEVA MD, 1993, J BIOL CHEM, V268, P19669