Presented at RNA2005, the Tenth Annual Meeting of the RNA Society, Banff, Alberta, Canada, May 24-29, 2005, and published in "RNA2005", p. 279, The RNA Society, Bethesda, MD 20814-3998, (2005).

"Sense and Antisense during RNA Initiation of the DNA Transcription Bubble".

Jeannette A. Hovsepian 1, @ and John H. Frenster 2, @

Departments of 1 Radiology and 2 Medicine, Stanford University School of Medicine, Stanford, California 94035, USA
@ Present Addresses: RNA Research, Physicians' Educational Series, Atherton, CA 94027-5446, USA
Phone:  650/367-6483;   FAX:   650/364-1773,  e-mail:   frensasc@ix.netcom.com
Supported in part by a USPHS Research Career Development Award CA-17857 to J.H.F.


Abstract:

The DNA Strand-Separation model of mammalian gene regulation (1) is based on the ability of DNA and RNA sequences to interact during the initiation and elongation phases of selective DNA transcription. RNA-DNA helices are more stable than DNA-DNA helices, and RNA-RNA helices are still more stable (2). Activator RNA is capable of binding to complementary DNA sequences in the anti-template DNA strand, and by such binding is capable of opening the DNA-DNA helix at selective sites for the initiation of DNA transcription on the DNA-template strand. As premessenger RNA is synthesized on the DNA-template strand, RNA splicing results in the formation of excised RNA exons, RNA introns, and RNA 5' leader sequences (3). Such excised 5' leader RNA sequences are the complement of the activator RNA sequences that initiated the transcription process. When DNA transcription is excessive at a particular gene locus, rising levels of messenger RNA and of 5' leader RNA from that locus are produced by the splicing process. Such increased levels of 5' leader RNA, specific for the given gene locus, are now capable of binding to activator RNA at the gene locus. Since RNA-RNA helices are more stable than DNA-RNA helices, activator RNA may be removed from the anti-template DNA strand at the gene locus. Such loss of activator RNA from the gene locus may result in a decrease of DNA transcription at that locus, thus providing a feedback-loop for the control of RNA synthesis at a particular gene locus. The RNA-RNA complex formed by such feedback may be very stable, and may be capable of storage during oogenesis, passage to daughter cells during mitosis, or transport to other nearby cells during embryonic induction (4).

Figure 1:
RNA-Induced Chromatin Remodeling and DNA Melting during Selective Gene Transcription (1, 2).

Figure 1:
Small nuclear RNA antisense species function as de-repressors of transcription by displacing repressor proteins (dark blocks), and then binding to the DNA anti-template strand at an initiation site (Fig. 1). This initiation stage frees the DNA template strand for transcription to gene-specific pre-messenger sense RNA, following the recruitment to that site, of RNA polymerase II and other transcription factors such as TFIIH.

     A specific de-repressor RNA sequence may interact with complementary anti-template DNA sequences at several gene loci, permitting one anti-sense RNA sequence population to activate multiple genes synchronously.




Figure 2:
Sense and Antisense during RNA Initiation of the DNA Transcription Bubble (3).

     An excessive synthesis of gene-specific  pre-messenger sense RNA may result in formation of RNA-RNA duplexes between the anti-sense activator RNA and the 5’ leader sequences of that pre-messenger sense RNA, removing the activator RNA from that initiation site, and reducing the pre-messenger RNA synthesis at that site in a feedback mechanism controlling selective gene dosage.

Activator RNA binds to the anti-template strand of the selected DNA locus, permitting transcription of gene-specific pre-messenger RNA (Fig. 2). The activator RNA of a given locus is complementary in base sequence to the 5' leader portion of the pre-messenger product. The pre-messenger RNA is spliced to form messenger RNA and 5' leader RNA . Splicing can occur directly or after formation of an inactive duplex RNA by base-pairing of activator RNA with the 5' leader. By such duplex formation, activator RNA may be selectively removed from the DNA locus, thus providing feedback inhibition of transcription of the DNA locus. With continued consumption of messenger RNA and degradation of 5' leader RNA, activator RNA may then be released from the inactive duplex, thus providing a positive feedback activation of transcription of the DNA locus. Different coding and non-coding genes may share 5' leaders with common base sequences, and thus be equally sensitive to a given species of activator RNA, both during selective gene transcription and during its selective inhibition (3).
 



References:

1. Frenster JH, "Mechanisms of Repression and De-Repression within Interphase Chromatin", In Vitro, vol. 1, pp. 78-101 (1965).

2. Frenster JH, "Correlation of the Binding to DNA Loops or to DNA Helices with the Effect on RNA Synthesis", Nature vol. 208, no. 5015, p. 1093 (December 11, 1965).

3. Herstein PR, and Frenster JH, "Mated Models of Gene Regulation in Eukaryotes", in: "Embryonic and Fetal Antigens in Cancer", vol. 2, pp. 5-7, Oak Ridge National Lab., Oak Ridge, Tenn., 1972.

4. Czihak G, "Evidence for Inductive Properties of the Micromere-RNA in Sea-urchin Embryos", Naturwissenschaften, vol. 52, no. 6, pp. 141-142 (1965).

5. Jabri E, "Non-Coding RNA: Small, but in Control", Nature Reviews Molecular Cell Biology, vol.6, no. 5, 361 (May, 2005).

6. Dell H, "Tumorigenesis: Micromanager", Nature Reviews Molecular Cell Biology, vol. 6, no. 5, 360, (May, 2005).

7. Johnson SM, Grosshans H, Shingara J, Byrom M, Jarvis R, Cheng A, Labourier E, Reinert KL, Brown D, and Slack FJ, "RAS Is Regulated by the let-7 MicroRNA Family".

8. Grosshans H, Johnson T, Reinert KL, Gerstein M, and Slack FJ, "The Temporal Patterning MicroRNA let-7 Regulates Several Transcription Factors at the Larval to Adult Transition in C. elegans".

9. Kuwabara T, Hsieh J, Nakashima K, Taira K, and Gage FH, "A Small Modulatory dsRNA Specifies the Fate of Adult Neural Stem Cells".

10. Muotri AR, Chu VT, Marchetto MCN, Deng W, Moran JV, and Gage FH, "Somatic mosaicism in neuronal precursor cells mediated by L1 retrotransposition".

11. Song X, Sun Y, and Garen A, "Roles of PSF protein and VL30 RNA in reversible gene regulation".


Links to RNA and Biological Causality:



Further Topics in:  Euchromatin,  active DNA, and  RNA  ribo-regulators:

Links to Euchromatin Activator RNA Reviews:
Links to Euchromatin Activator RNA Research:
Links to Ultrastructural Probes of DNase I-Sensitive Sites:
Links to RNA as a Therapeutic Agent:
Links to Hodgkin Lymphoma Immuno-Pathology:
Links to Activated T-Lymphocyte Immunotherapy:
Links to Medical Systems Biology:
Links to Selective Gene Transcription:
Links to RNA-Induced Epigenetics:
Links to RNA-Induced Embryogenesis:
Links to RNA and Biological Causality:
Links to Reprogramming and Neoplasia:

A Brief History of Activator RNA:

"Ultrastructural Probes of Active DNA Sites, and the RNA Activators of DNA". (PowerPoint Presentation).




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