Diverse roles of hnRNP L in mammalian mRNA processing: a combined microarray and RNAi analysis.
Alternative mRNA splicing patterns are determined by the combinatorial control of regulator proteins and their target RNA sequences. We have recently characterized human hnRNP L as a global regulator of alternative splicing, binding to diverse C/A-rich elements. To systematically identify hnRNP L target genes on a genome-wide level, we have combined splice-sensitive microarray analysis and an RNAi-knockdown approach. As a result, we describe 11 target genes of hnRNP L that were validated by RT-PCR and that represent several new modes of hnRNP L-dependent splicing regulation, involving both activator and repressor functions: first, intron retention; second, inclusion or skipping of cassette-type exons; third, suppression of multiple exons; and fourth, alternative poly(A) site selection. In sum, this approach revealed a surprising diversity of splicing-regulatory processes as well as poly(A) site selection in which hnRNP L is involved.
Hung
Lee-Hsueh
LH
Institute of Biochemistry, Justus-Liebig-University of Giessen, D-35392 Giessen, Germany.
Heiner
Monika
M
Hui
Jingyi
J
Schreiner
Silke
S
Benes
Vladimir
V
Bindereif
Albrecht
A
D017398:Alternative Splicing / Q000235:genetics*; D020816:Amino Acid Motifs; D005091:Exons; D006367:HeLa Cells; D034664:Heterogeneous-Nuclear Ribonucleoprotein L / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D006801:Humans; D007438:Introns; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D026723:Polyadenylation; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism*
D034664:Heterogeneous-Nuclear Ribonucleoprotein L; D012333:RNA, Messenger