Functional pleiotropy of an intramolecular triplex-forming fragment from the 3'-UTR of the rat Pigr gene.

I. Fabregat, K. S. Koch, T. Aoki, A. E. Atkinson, H. Dang, O. Amosova, J. R. Fresco, Carl L. Schildkraut, H. L. Leffert

Research output: Contribution to journalArticle

Abstract

A microsatellite-containing 359-bp restriction fragment, isolated from the rat Pigr gene (murine polymeric immunoglobulin receptor gene) 3'-untranslated region (3'-UTR) and inserted into 3'-UTR or 3' flanking positions in transcription units of supercoiled plasmids, attenuates luciferase reporter gene expression in orientation- and position-dependent ways following transient transfection of human 293 cells. The same fragment stimulates orientation-dependent gene expression in a 5' flanking position. Plasmid linearization abrogates both orientation- and position-dependent responses. Cell-free translation reveals that 5' and 3' flanking expression responses are proportional to increased and decreased luciferase mRNA levels, whereas 3'-UTR expression is associated with control mRNA levels. Hypersensitivity to nucleases S1 and P1, gel mobility differences between supercoiled plasmids carrying opposing microsatellite orientations, and anomalous melting profiles of this fragment are also observed. These results suggest that functional pleiotropy of this fragment depends on the DNA context of its purine-rich microsatellite strand and on DNA supercoiling. Intramolecular triplexes stabilized by supercoiling and secondary structures of purine repeat-rich mRNAs may also confer regulatory properties to similar genomic elements.

Original languageEnglish (US)
Pages (from-to)53-65
Number of pages13
JournalPhysiol Genomics
Volume5
Issue number2
StatePublished - Mar 8 2001
Externally publishedYes

Fingerprint

3' Untranslated Regions
Microsatellite Repeats
Plasmids
Luciferases
Messenger RNA
Polymeric Immunoglobulin Receptors
Genes
Gene Expression
Immunoglobulin Genes
DNA
Reporter Genes
Freezing
Transfection
Hypersensitivity
Gels
purine

ASJC Scopus subject areas

  • Physiology
  • Genetics

Cite this

Fabregat, I., Koch, K. S., Aoki, T., Atkinson, A. E., Dang, H., Amosova, O., ... Leffert, H. L. (2001). Functional pleiotropy of an intramolecular triplex-forming fragment from the 3'-UTR of the rat Pigr gene. Physiol Genomics, 5(2), 53-65.

Functional pleiotropy of an intramolecular triplex-forming fragment from the 3'-UTR of the rat Pigr gene. / Fabregat, I.; Koch, K. S.; Aoki, T.; Atkinson, A. E.; Dang, H.; Amosova, O.; Fresco, J. R.; Schildkraut, Carl L.; Leffert, H. L.

In: Physiol Genomics, Vol. 5, No. 2, 08.03.2001, p. 53-65.

Research output: Contribution to journalArticle

Fabregat, I, Koch, KS, Aoki, T, Atkinson, AE, Dang, H, Amosova, O, Fresco, JR, Schildkraut, CL & Leffert, HL 2001, 'Functional pleiotropy of an intramolecular triplex-forming fragment from the 3'-UTR of the rat Pigr gene.', Physiol Genomics, vol. 5, no. 2, pp. 53-65.
Fabregat I, Koch KS, Aoki T, Atkinson AE, Dang H, Amosova O et al. Functional pleiotropy of an intramolecular triplex-forming fragment from the 3'-UTR of the rat Pigr gene. Physiol Genomics. 2001 Mar 8;5(2):53-65.
Fabregat, I. ; Koch, K. S. ; Aoki, T. ; Atkinson, A. E. ; Dang, H. ; Amosova, O. ; Fresco, J. R. ; Schildkraut, Carl L. ; Leffert, H. L. / Functional pleiotropy of an intramolecular triplex-forming fragment from the 3'-UTR of the rat Pigr gene. In: Physiol Genomics. 2001 ; Vol. 5, No. 2. pp. 53-65.
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