Conserved zinc fingers mediate multiple functions of ZFP100, a U7snRNP associated protein.
Formation of the 3' end of replication-dependent histone mRNAs is most robust during S phase and is mediated by both the stem-loop binding protein (SLBP) and the U7 snRNP. We previously identified a 100-kDa zinc finger protein (ZFP100) as a component of U7 snRNP that interacts with the SLBP/pre-mRNA complex. Here, we show that myc- or GFP-tagged ZFP100 overexpressed after transfection is concentrated in Cajal bodies (CBs), and unlike components of the spliceosomal snRNPs, photobleaching experiments demonstrate that ZFP100 is stably associated with CBs. Of the 18 zinc fingers contained within ZFP100, the region encompassing fingers 2-6 is sufficient to maintain CB localization. Zn fingers 5-10 are required for maximal binding of ZFP100 to a 20-amino-acid region of Lsm11, a U7 snRNP core protein. Expression of ZFP100 stimulates histone mRNA processing in vivo, assayed by activation of a reporter gene that encodes a GFP mRNA ending in a histone 3' end. Importantly, the domain that is required for CB localization and Lsm11 binding is also sufficient to stimulate histone pre-mRNA processing in vivo. Comparisons with other mammalian ZFP100 orthologs show that the central Zn fingers sufficient for in vivo activity are most highly conserved, whereas the number and sequence of the Zn fingers in the N- and C-terminal domains vary.
Wagner
Eric J
EJ
Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
Ospina
Jason K
JK
Hu
Yue
Y
Dundr
Miroslav
M
Matera
A Gregory
AG
Marzluff
William F
WF
D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D020541:Coiled Bodies / Q000648:ultrastructure; D017124:Conserved Sequence; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics*; D005434:Flow Cytometry; D017353:Gene Deletion; D006367:HeLa Cells; D006657:Histones / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids; D020885:Ribonucleoprotein, U7 Small Nuclear / Q000737:chemistry* / Q000235:genetics; D016335:Zinc Fingers
D004268:DNA-Binding Proteins; D006657:Histones; D020885:Ribonucleoprotein, U7 Small Nuclear; C095988:ZNF473 protein, human