A. Wang et al., IDENTIFICATION AND CHARACTERIZATION OF HUMAN GENES ENCODING HPRP3P AND HPRP4P, INTERACTING COMPONENTS OF THE SPLICEOSOME, Human molecular genetics, 6(12), 1997, pp. 2117-2126
Nuclear RNA splicing occurs in an RNA-protein complex, termed the spli
ceosome. U4/U6 snRNP is one of four essential small nuclear ribonucleo
protein (snRNP) particles (U1, U2, U5 and U4/U6) present in the splice
osome. U4/U6 snRNP contains two snRNAs (U4 and U6) and a number of pro
teins, We report here the identification and characterization of two h
uman genes encoding U4/U6-associated splicing factors, Hprp3p and Hprp
4p, respectively. Hprp3p is a 77 kDa protein, which is homologous to t
he Saccharomyces cerevisiae splicing factor Prp3p, Amino acid sequence
analysis revealed two putative homologues in Caenorhabditis elegans a
nd Schizosaccharomyces pombe. Polyclonal antibodies against Hprp3p wer
e generated with His-tagged Hprp3p over-produced in Escherichia coli,
This splicing factor can co-immunoprecipitate with U4, U6 and U5 snRNA
s, suggesting that it is present in the U4/U6 U5 tri-snRNP, Hprp4p is
a 58 kDa protein homologous to yeast splicing factor Prp4p, Like yeast
Prp4p, the human homologue contains repeats homologous to the beta-su
bunit of G-proteins, These repeats are called WD repeats because there
is a highly conserved dipeptide of tryptophan and aspartic acid prese
nt at the end of each repeat, The primary amino acid sequence homology
between human Hprp4p and yeast Prp4p led to the discovery of two addi
tional WD repeats in yeast Prp4p, Structural homology between these hu
man and yeast splicing factors and the beta-subunit of G-proteins has
been identified by sequence-similarity comparison and analysis of the
protein folding by threading, Structural models of Hprp4p and Prp4p wi
th a seven-blade beta-propeller topology have been generated based on
the structure of beta-transducin, Hprp3p and Hprp4p have been shown to
interact with each other and the first 100 amino acids of Hprp3p are
not essential for this interaction, These experiments suggest that bot
h Hprp3p and Hprp4p are components of human spliceosomes.