Network of interactions of a novel plant-specific Arg/Ser-rich protein, atRSZ33, with atSC35-like splicing factors

被引:70
作者
Lopato, S [1 ]
Forstner, C [1 ]
Kalyna, M [1 ]
Hilscher, J [1 ]
Langhammer, U [1 ]
Indrapichate, K [1 ]
Lorkovic, ZJ [1 ]
Barta, A [1 ]
机构
[1] Univ Vienna, Vienna Bioctr, Inst Med Biochem, A-1030 Vienna, Austria
关键词
D O I
10.1074/jbc.M206455200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Arg/Ser-rich (RS) proteins play a crucial role in splicing and are implicated in splice site selection in metazoa. In plants, intron recognition seems to differ from the one in animals due to specific factor requirements. Here we describe a new plant-specific RS-rich protein, atRSZ33, with a unique domain structure consisting of an RNA recognition motif (RRM), two zinc knuckles embedded in a basic RS region, and an acidic C-terminal domain. atRSZ33 was found to be a phosphoprotein that concentrates in nuclear speckles and is predominantly present in roots and flowers. In a yeast two-hybrid screen, atRSZ33 interacted with splicing factors atSRp34/SR1, an Arabidopsis ortholog of human SF2/ASF; atRSZp21 and atRSZp22, which are similar to the human 9G8; and three novel SC35-like splicing factors termed atSCL28, atSCL30, and atSCL33/SR33. Two further members of the SCL family, namely SCL30a and the ortholog of mammalian SC35, atSC35, were also found to interact with atRSZ33. These interactions were verified by in vitro binding assays; furthermore, the transcriptional activity of atRSZ33 was found to overlap with the ones of its interacting partners. These specific interactions coupled with the many similarities of atRSZ33 to SR proteins suggest that its main activity is in spliceosome assembly. Mapping of regions necessary for protein-protein interaction between atRSZ33 and atSCL33/SR33 revealed that both zinc knuckles together with a small part of the RS and the RRM domain are required for efficient binding. However, the interacting domain is relatively small, allowing binding of additional proteins, a feature that is consistent with the proposed role of atRSZ33 in spliceosome assembly.
引用
收藏
页码:39989 / 39998
页数:10
相关论文
共 65 条
[31]   atSRp30, one of two SF2/ASF-like proteins from Arabidopsis thaliana, regulates splicing of specific plant genes [J].
Lopato, S ;
Kalyna, M ;
Dorner, S ;
Kobayashi, R ;
Krainer, AR ;
Barta, A .
GENES & DEVELOPMENT, 1999, 13 (08) :987-1001
[32]   Characterization of a novel arginine/serine-rich splicing factor in Arabidopsis [J].
Lopato, S ;
Waigmann, E ;
Barta, A .
PLANT CELL, 1996, 8 (12) :2255-2264
[33]   RBP45 and RBP47, two oligouridylate-specific hnRNP-like proteins interacting with poly(A)+ RNA in nuclei of plant cells [J].
Lorkovic, ZJ ;
Kirk, DAW ;
Klahre, U ;
Hemmings-Mieszczak, M ;
Filipowicz, W .
RNA, 2000, 6 (11) :1610-1624
[34]   Pre-mRNA splicing in higher plants [J].
Lorkovic, ZJ ;
Kirk, DAW ;
Lambermon, MHL ;
Filipowicz, W .
TRENDS IN PLANT SCIENCE, 2000, 5 (04) :160-167
[35]   Genome analysis:: RNA recognition motif (RRM) and K homology (KH) domain RNA-binding proteins from the flowering plant Arabidopsis thaliana [J].
Lorkovic, ZJ ;
Barta, A .
NUCLEIC ACIDS RESEARCH, 2002, 30 (03) :623-635
[36]   SR proteins and splicing control [J].
Manley, JL ;
Tacke, R .
GENES & DEVELOPMENT, 1996, 10 (13) :1569-1579
[37]   Inhibition of msl-2 splicing by Sex-lethal reveals interaction between U2AF35 and the 3′ splice site AG [J].
Merendino, L ;
Guth, S ;
Bilbao, D ;
Martinez, C ;
Valcárcel, J .
NATURE, 1999, 402 (6763) :838-841
[38]   RNA splicing: What has phosphorylation got to do with it? [J].
Misteli, T .
CURRENT BIOLOGY, 1999, 9 (06) :R198-+
[39]   Serine phosphorylation of SR proteins is required for their recruitment to sites of transcription in vivo [J].
Misteli, T ;
Cáceres, JF ;
Clement, JQ ;
Krainer, AR ;
Wilkinson, MF ;
Spector, DL .
JOURNAL OF CELL BIOLOGY, 1998, 143 (02) :297-307
[40]   Protein phosphorylation and the nuclear organization of pre-mRNA splicing [J].
Misteli, T ;
Spector, DL .
TRENDS IN CELL BIOLOGY, 1997, 7 (04) :135-138