The fission yeast methyl phosphate capping enzyme Bmc1 guides 2′-O-methylation of the U6 snRNA

Abstract

Abstract Splicing requires the tight coordination of dynamic spliceosomal RNAs and proteins. U6 is the only spliceosomal RNA transcribed by RNA Polymerase III and undergoes an extensive maturation process. In humans and fission yeast, this includes addition of a 5′ γ-monomethyl phosphate cap by members of the Bin3/MePCE family as well as snoRNA guided 2′-O-methylation. Previously, we have shown that the Bin3/MePCE homolog Bmc1 is recruited to the S. pombe telomerase holoenzyme by the LARP7 family protein Pof8, where it acts in a catalytic-independent manner to protect the telomerase RNA and facilitate holoenzyme assembly. Here, we show that Bmc1 and Pof8 are required for the formation of a distinct U6 snRNP that promotes 2′-O-methylation of U6, and identify a non-canonical snoRNA that guides this methylation. We also show that the 5′ γ-monomethyl phosphate capping activity of Bmc1 is not required for its role in promoting snoRNA guided 2′-O-methylation, and that this role relies on different regions of Pof8 from those required for Pof8 function in telomerase. Our results are consistent with a novel role for Bmc1/MePCE family members in stimulating 2′-O-methylation and a more general role for Bmc1 and Pof8 in guiding noncoding RNP assembly beyond the telomerase RNP.