Relatively Stable Population of c-myc RNA that Lacks Long Poly(A)

Abstract

We examined the turnover of c-myc RNA in the human promyelocytic cell line HL-60. In whole-cell RNA from rapidly growing cells we observed two major size classes of c-myc RNA, 2.4 and 2.2 kilobases (kb). When HL-60 cells were treated with actinomycin D for 30 min to inhibit transcription, the 2.4-kb c-myc RNA population was rapidly degraded, while the 2.2-kb c-myc RNA was degraded much more slowly. S1 nuclease transcript mapping and promoter-specific probes were utilized to show that both the stable 2.2-kb and the labile 2.4-kb c-myc RNA populations have 5' ends at the second promoter site (P2) and 3' ends at the second poly(A) addition site. To examine further possible structural differences between these two RNA populations, we fractionated RNA on an oligo(dT)-cellulose column to separate RNAs that contained long poly(A) tails from those which did not. We found that the labile 2.4-kb c-myc RNA population bound to oligo(dT)-cellulose, while the more stable 2.2-kb c-myc RNA population did not. Preliminary estimates of their half-lives (t1/2) showed that the poly(A)+ c-myc RNA had a t1/2 of 12 min, while the c-myc RNA that did not bind to oligo(dT)-cellulose had a t1/2 of greater than 1 h. Several other cell types contain both poly(A)+ and nonpoly(A)+ c-myc RNAs including HeLa cells, normal human bone marrow cells, and normal mouse fetal liver cells. In agreement with the results in HL-60 cell, HeLa cell poly(A)+ c-myc RNA was more labile than c-myc RNA that lacked poly(A). The stable, nonpoly(A)+ c-myc RNA population may be important in the posttranscriptional regulation of c-myc expression.