OsRPA2

Annotated Information

===Function

Replication protein A (RPA) is a heterotrimeric, singlestranded DNA-binding protein with several functions in DNA metabolism in humans and yeast and supposedly also in plants. OsRPA2, the 32-kDa subunit of RPA from rice (Oryza sativa L.).The plant-specific mechanisms of regulation have evolved for RPA2 within the generally well-conserved process of DNA replication, suggesting specific requirements for regulation of DNA metabolism in plants as compared to other eukaryotes.

Expression

Replication protein A (RPA) is a heterotrimeric protein complex, encoded by RPA1, RPA2 and RPA3.The respective proteins in humans were termed RPA70,RPA32 and RPA14 according to their molecular weights. Even though the precise function of RPA is still not clear, extensive, mostly biochemical analysis of RPA proteins in humans and yeast led to the conclusion that RPA plays essential roles in many aspects of nucleic acid metabolism, including DNA replication, but also DNA repair and DNA recombination .RPA binds single-stranded DNA during replication to stabilize DNA in the single-strand conformation(Treuner et al. 1996). It stimulates DNA synthesisthrough interaction with DNA polymerase a (Kennyet al. 1990). Mutations in RPA2 were found to be lethal in yeast(Philipova et al. 1996). The RPA32 protein isphosphorylated at the G1- to S-phase transition (Fang and Newport 1993) and is hyperphosphorylated after DNA damage. Hyperphosphorylation of RPA32 possibly results in disassembly of the trimeric complex(Treuner et al. 1999 a, 1999b).Much less is known about RPA from plants. An RPA protein complex was isolated from tobacco by binding and elution from single-stranded DNA and was shown to induce DNA polymerase activity (Garcia-Maya and Buck 1997). From rice, cDNAs of the two subunits OsRPA1 (van der Knaap et al. 1997; Ishibashi et al.2001) and OsRPA2 (Ishibashi et al. 2001; this study) were isolated. The OsRPA1 gene was shown to be expressed mainly in proliferative tissues and was induced coordinately with cell cycle induction by gibberellic acid(van der Knaap 1997) and during re-growth after starvation of suspension cells (Ishibashi et al. 2001). A gene,coding for the RPA3 subunit has not been identified from plants to date.We describe here, the gene and protein regulation of OsRPA2 from rice. The results presented indicate that despite a high degree of conservation of RPA among eukaryotes, regulation of OsRPA2 displays features that appear to be specific to plants

Evolution

A partial cDNA of 335 nt comprising the 3¢-untranslated region(UTR) and 80 nt of the coding sequence at the C-terminus was isolated by subtractive hybridization as described by Lorbiecke and Sauter (2002). Through PCR from a rice cDNA library (Sauter et al. 1995) using a gene-specific primer (bp 896–915) and a plasmid-derived primer (bp 798–817 of pBluescript II SK) ,a longer clone was obtained that included the coding region up to the start codon. To obtain the 5¢-UTR of the cDNA, a 5¢-rapid amplification of cDNA ends(5¢-RACE; Gibco, Karlsruhe,Germany) was performed using cDNA as template that was reverse-transcribed from total RNA isolated from the intercalary meristem of plants that were submerged for 6 h. The complete sequence with the encoded open reading frame was termed OsRPA2and was submitted to the nucleotide sequence database at EMBL (accession number AJ278822).

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Labs working on this gene

Department of Applied Biological Science,Faculty of Science and Technology, Science University of Tokyo, 2641 Yamazaki,278-8510 Noda-shi, Chiba-ken, Tokyo, Japan

References

[1] Plant-specific regulation of replication protein A2 (OsRPA2) from rice during the cell cycle and in response to ultraviolet light exposure

 Planta, 2003, 217(3): 457-465.

[2]Wold MS (1997) Replication protein A: a heterotrimeric, singlestranded DNA-binding protein required for eukaryotic DNA metabolism. Annu Rev Biochem 66:61–92.

[3]Treuner K, Ramsperger U, Knippers R (1996) Replication protein A induces the unwinding of long double-stranded DNA regions. J Mol Biol 259:104–112.

Structured Information

Gene Name	{{{GeneName}}}
Description	{{{Description}}}
Version	{{{Version}}}
Length	{{{Length}}}
Definition	{{{Definition}}}
Source	{{{Source}}}
Chromosome	Chromosome 2
Location	Chromosome 2:36526606..36529420
Sequence Coding Region	36526967..36527051,36527165..36527233,36527599..36527681,36527762..36527857,36527983..36528061 ,36528143..36528220,36528357..36528398,36528803..36528900,36528996..36529115 ,36529208..36529297
Expression	GEO Profiles:{{{GeneName}}}
Genome Context	<gbrowseImage1> name=OsRPA2 source=RiceChromosome02 preset=GeneLocation </gbrowseImage1>
Gene Structure	<gbrowseImage2> name=OsRPA2 source=RiceChromosome02 preset=GeneLocation </gbrowseImage2>
Coding Sequence	AA = <aaseq>MMSFSQPDAFSPSQFTSSQNAAADSTTPSKSRGASSTMPLTVKQ ISEAQQSGITGEKGAPFVVDGVETANVRLVGLVSGKTERNTDVSFTIDDGTGRLDFIR WVNDGADSAETAAVQNGMYVSVIGSLKGLQERKRATAFAIRPVTDYNEVTLHFIQCVR MHLENTKSQIGSPAKTYSAMGSSSSNGFSEMTTPTSVKSNPAPVLSVTNGSKTDLNTE VLNVFREPANVESEHGVHIDEIVKRFRLPEAKIKVAIDYLADIGHIYSTIDESHYKSA FNE</aaseq>
Protein Sequence	{{{AA}}}
Gene Sequence	Link = http://www.ricedata.cn/gene/list/1988.htm
External Link(s)	NCBI Gene:{{{GeneName}}}, {{{Link}}}