RNA-Directed Actions of 8-Chloro-Adenosine in Multiple Myeloma Cells

Christine M. Stellrecht, Carlos O. Rodriguez, Mary Ayres, Varsha Gandhi

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

The purine analogue, 8-chloro-adenosine (8-Cl-Ado), induces apoptosis in a number of multiple myeloma (MM) cell lines. This ribonucleoside analogue accumulates as a triphosphate and selectively inhibits RNA synthesis without perturbing DNA synthesis. Cellular RNA is synthesized by one of three polymerases (Pol I, II, or III); thus, the inhibition of one or more RNA polymerases may be mediating 8-Cl-Ado cytotoxicity. Here, we have addressed this question by dissecting the RNA-directed actions of 8-Cl-Ado in MM cells. Differential alterations in [3H]uridine incorporation were found in the three major classes of RNA after a 20-h exposure with 10 μM 8-Cl-Ado. The synthesis rate of Pol III transcripts, 5 S and tRNA, remained unchanged, whereas Pol I-mediated rRNA synthesis decreased by ∼20%. In contrast, mRNA synthesis, which is transcribed by Pol II, rapidly declined within 4 h and reached a 50% decrease, which was maintained for 20 h. Parallel to RNA synthesis inhibition, 8-Cl-Ado was maximally incorporated in the mRNA (>13 nmol/mg RNA), which was 5-fold higher than the tRNA and rRNA incorporation. Electrophoretic and radiographic analysis of newly synthesized and processed [14C]uridine-labeled transcripts indicated that the analogue blocks transcription elongation. Consistent with that result, high-performance liquid chromatography analysis of micrococcal nuclease and spleen phosphodi-esterase-digested RNA demonstrated that the analogue incorporation is at the 3′ terminus. In conclusion, our data demonstrate that in MM cells, 8-Cl-Ado is preferentially incorporated into mRNA, suggesting a propensity toward Pol II, and inhibits RNA synthesis by premature transcriptional chain termination.

Original languageEnglish (US)
Pages (from-to)7968-7974
Number of pages7
JournalCancer Research
Volume63
Issue number22
StatePublished - Nov 15 2003
Externally publishedYes

Fingerprint

Multiple Myeloma
Adenosine
RNA
Uridine
Transfer RNA
Messenger RNA
Ribonucleosides
Micrococcal Nuclease
RNA Polymerase II
Esterases
DNA-Directed RNA Polymerases
Spleen
High Pressure Liquid Chromatography
Apoptosis
Cell Line
DNA

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Stellrecht, C. M., Rodriguez, C. O., Ayres, M., & Gandhi, V. (2003). RNA-Directed Actions of 8-Chloro-Adenosine in Multiple Myeloma Cells. Cancer Research, 63(22), 7968-7974.

RNA-Directed Actions of 8-Chloro-Adenosine in Multiple Myeloma Cells. / Stellrecht, Christine M.; Rodriguez, Carlos O.; Ayres, Mary; Gandhi, Varsha.

In: Cancer Research, Vol. 63, No. 22, 15.11.2003, p. 7968-7974.

Research output: Contribution to journalArticle

Stellrecht, CM, Rodriguez, CO, Ayres, M & Gandhi, V 2003, 'RNA-Directed Actions of 8-Chloro-Adenosine in Multiple Myeloma Cells', Cancer Research, vol. 63, no. 22, pp. 7968-7974.
Stellrecht CM, Rodriguez CO, Ayres M, Gandhi V. RNA-Directed Actions of 8-Chloro-Adenosine in Multiple Myeloma Cells. Cancer Research. 2003 Nov 15;63(22):7968-7974.
Stellrecht, Christine M. ; Rodriguez, Carlos O. ; Ayres, Mary ; Gandhi, Varsha. / RNA-Directed Actions of 8-Chloro-Adenosine in Multiple Myeloma Cells. In: Cancer Research. 2003 ; Vol. 63, No. 22. pp. 7968-7974.
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