Translation Regulation and Proteasome Mediated Degradation Cooperate to Keep Stem-Loop Binding Protein Low in G1-Phase: REGULATION OF SLBP EXPRESSION IN G1-PHASE

Histone mRNA levels are cell cycle regulated, and the major regulatory steps are at the posttranscriptional level. A major regulatory mechanism is S- phase restriction of Stem-loop binding protein (SLBP) which binds to the 3′ end of histone mRNA and participates in multiple steps of histone mRNA metabolism, including 3′ end processing, translation and regulation of mRNA stability. SLBP expression is cell cycle regulated without significant change in its mRNA level. SLBP expression is low in G1 until just before S phase where it functions and at the end of S phase SLBP is degraded by proteasome complex depending on phosphorylations on Thr 60 and Thr61. Here using synchronized HeLa cells we showed that SLBP production rate is low in early G1 and recovers back to S phase level somewhere between early and mid-G1. Further, we showed that SLBP is unstable in G1 due to proteasome mediated degradation as a novel mechanism to keep SLBP low in G1. Finally, the S/G2 stable mutant form of SLBP is degraded by proteasome in G1, indicating that the SLBP degradation mechanism in G1 is independent of the previously identified S/G2 degradation mechanism. In conclusion, as a mechanism to limit histone production to S phase, SLBP is kept low in G1 phase due to cooperative action of translation regulation and proteasome mediated degradation which is independent of previously known S/G2 degradation