UniProt functional annotation for P15919



	UniProt functional annotation for P15919

UniProt code: P15919.

Organism: Mus musculus (Mouse).

Taxonomy: Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia; Eutheria; Euarchontoglires; Glires; Rodentia; Myomorpha; Muroidea; Muridae; Murinae; Mus; Mus.

Function: Catalytic component of the RAG complex, a multiprotein complex that mediates the DNA cleavage phase during V(D)J recombination. V(D)J recombination assembles a diverse repertoire of immunoglobulin and T-cell receptor genes in developing B and T- lymphocytes through rearrangement of different V (variable), in some cases D (diversity), and J (joining) gene segments. In the RAG complex, RAG1 mediates the DNA-binding to the conserved recombination signal sequences (RSS) and catalyzes the DNA cleavage activities by introducing a double-strand break between the RSS and the adjacent coding segment. RAG2 is not a catalytic component but is required for all known catalytic activities. DNA cleavage occurs in 2 steps: a first nick is introduced in the top strand immediately upstream of the heptamer, generating a 3'-hydroxyl group that can attack the phosphodiester bond on the opposite strand in a direct transesterification reaction, thereby creating 4 DNA ends: 2 hairpin coding ends and 2 blunt, 5'-phosphorylated ends. The chromatin structure plays an essential role in the V(D)J recombination reactions and the presence of histone H3 trimethylated at 'Lys-4' (H3K4me3) stimulates both the nicking and haipinning steps. The RAG complex also plays a role in pre-B cell allelic exclusion, a process leading to expression of a single immunoglobulin heavy chain allele to enforce clonality and monospecific recognition by the B-cell antigen receptor (BCR) expressed on individual B-lymphocytes. The introduction of DNA breaks by the RAG complex on one immunoglobulin allele induces ATM- dependent repositioning of the other allele to pericentromeric heterochromatin, preventing accessibility to the RAG complex and recombination of the second allele. In addition to its endonuclease activity, RAG1 also acts as an E3 ubiquitin-protein ligase that mediates monoubiquitination of histone H3. Histone H3 monoubiquitination is required for the joining step of V(D)J recombination. Mediates polyubiquitination of KPNA1. {ECO:0000269|PubMed:10601032, ECO:0000269|PubMed:10678172, ECO:0000269|PubMed:12629039, ECO:0000269|PubMed:14671314, ECO:0000269|PubMed:17028591, ECO:0000269|PubMed:19118899, ECO:0000269|PubMed:19396172, ECO:0000269|PubMed:19448632, ECO:0000269|PubMed:19524534, ECO:0000269|PubMed:20122409, ECO:0000269|PubMed:22157821, ECO:0000269|PubMed:2598259, ECO:0000269|PubMed:8521468, ECO:0000269|PubMed:9094713}.

Catalytic activity: Reaction=S-ubiquitinyl-[E2 ubiquitin-conjugating enzyme]-L-cysteine + [acceptor protein]-L-lysine = [E2 ubiquitin-conjugating enzyme]-L- cysteine + N(6)-ubiquitinyl-[acceptor protein]-L-lysine.; EC=2.3.2.27;

Cofactor: Name=Mg(2+); Xref=ChEBI:CHEBI:18420; Evidence={ECO:0000269|PubMed:10601032}; Name=Mn(2+); Xref=ChEBI:CHEBI:29035; Evidence={ECO:0000269|PubMed:10601032}; Note=Binds 1 divalent metal cation per subunit. Mg(2+) or Mn(2+). {ECO:0000269|PubMed:10601032};

Subunit: Homodimer. Component of the RAG complex composed of core components RAG1 and RAG2, and associated component HMGB1 or HMGB2. Interacts with DCAF1, leading to recruitment of the CUL4A-RBX1-DDB1- DCAF1/VPRBP complex to ubiquitinate proteins and limit error-prone repair during V(D)J recombination. {ECO:0000269|PubMed:19396172, ECO:0000269|PubMed:22157821, ECO:0000269|PubMed:8521468, ECO:0000269|PubMed:9094713, ECO:0000269|PubMed:9184213, ECO:0000269|PubMed:9228952}.

Subcellular location: Nucleus {ECO:0000255|PROSITE-ProRule:PRU00820, ECO:0000269|PubMed:8284210}.

Tissue specificity: Maturing lymphoid cells and central nervous system.

Domain: The RING-type zinc finger mediates the E3 ubiquitin-protein ligase activity. {ECO:0000269|PubMed:19396172}.

Domain: The NBD (nonamer binding) DNA-binding domain mediates the specific binding to the nonamer RSS motif by forming a tightly interwoven homodimer that binds and synapses 2 nonamer elements, with each NBD making contact with both DNA molecules. Each RSS is composed of well-conserved heptamer (consensus 5'-CACAGTG-3') and nonamer (consensus 5'-ACAAAAACC-3') sequences separated by a spacer of either 12 bp or 23 bp. {ECO:0000255|PROSITE-ProRule:PRU00820, ECO:0000269|PubMed:19396172}.

Ptm: Autoubiquitinated in the presence of CDC34/UBCH3. {ECO:0000269|PubMed:14671314, ECO:0000269|PubMed:19118899}.

Disruption phenotype: Mice display a severe combined immunodeficiency phenotype. The have a small lymphoid organs that do not contain mature B and T-lymphocytes. The arrest of B- and T-cell differentiation occurs at an early stage and correlates with the inability to perform V(D)J recombination. The frequency of homologous immunoglobulin pairing is much lower. No obvious neuroanatomical or behavioral abnormalities have been observed. {ECO:0000269|PubMed:1547488}.

Similarity: Belongs to the RAG1 family. {ECO:0000255|PROSITE- ProRule:PRU00820}.

Annotations taken from UniProtKB at the EBI.


Organism:		Mus musculus (Mouse).
Taxonomy:		Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia; Eutheria; Euarchontoglires; Glires; Rodentia; Myomorpha; Muroidea; Muridae; Murinae; Mus; Mus.



Function:		Catalytic component of the RAG complex, a multiprotein complex that mediates the DNA cleavage phase during V(D)J recombination. V(D)J recombination assembles a diverse repertoire of immunoglobulin and T-cell receptor genes in developing B and T- lymphocytes through rearrangement of different V (variable), in some cases D (diversity), and J (joining) gene segments. In the RAG complex, RAG1 mediates the DNA-binding to the conserved recombination signal sequences (RSS) and catalyzes the DNA cleavage activities by introducing a double-strand break between the RSS and the adjacent coding segment. RAG2 is not a catalytic component but is required for all known catalytic activities. DNA cleavage occurs in 2 steps: a first nick is introduced in the top strand immediately upstream of the heptamer, generating a 3'-hydroxyl group that can attack the phosphodiester bond on the opposite strand in a direct transesterification reaction, thereby creating 4 DNA ends: 2 hairpin coding ends and 2 blunt, 5'-phosphorylated ends. The chromatin structure plays an essential role in the V(D)J recombination reactions and the presence of histone H3 trimethylated at 'Lys-4' (H3K4me3) stimulates both the nicking and haipinning steps. The RAG complex also plays a role in pre-B cell allelic exclusion, a process leading to expression of a single immunoglobulin heavy chain allele to enforce clonality and monospecific recognition by the B-cell antigen receptor (BCR) expressed on individual B-lymphocytes. The introduction of DNA breaks by the RAG complex on one immunoglobulin allele induces ATM- dependent repositioning of the other allele to pericentromeric heterochromatin, preventing accessibility to the RAG complex and recombination of the second allele. In addition to its endonuclease activity, RAG1 also acts as an E3 ubiquitin-protein ligase that mediates monoubiquitination of histone H3. Histone H3 monoubiquitination is required for the joining step of V(D)J recombination. Mediates polyubiquitination of KPNA1. {ECO:0000269\|PubMed:10601032, ECO:0000269\|PubMed:10678172, ECO:0000269\|PubMed:12629039, ECO:0000269\|PubMed:14671314, ECO:0000269\|PubMed:17028591, ECO:0000269\|PubMed:19118899, ECO:0000269\|PubMed:19396172, ECO:0000269\|PubMed:19448632, ECO:0000269\|PubMed:19524534, ECO:0000269\|PubMed:20122409, ECO:0000269\|PubMed:22157821, ECO:0000269\|PubMed:2598259, ECO:0000269\|PubMed:8521468, ECO:0000269\|PubMed:9094713}.


Catalytic activity:		Reaction=S-ubiquitinyl-[E2 ubiquitin-conjugating enzyme]-L-cysteine + [acceptor protein]-L-lysine = [E2 ubiquitin-conjugating enzyme]-L- cysteine + N(6)-ubiquitinyl-[acceptor protein]-L-lysine.; EC=2.3.2.27;
Cofactor:		Name=Mg(2+); Xref=ChEBI:CHEBI:18420; Evidence={ECO:0000269\|PubMed:10601032}; Name=Mn(2+); Xref=ChEBI:CHEBI:29035; Evidence={ECO:0000269\|PubMed:10601032}; Note=Binds 1 divalent metal cation per subunit. Mg(2+) or Mn(2+). {ECO:0000269\|PubMed:10601032};
Subunit:		Homodimer. Component of the RAG complex composed of core components RAG1 and RAG2, and associated component HMGB1 or HMGB2. Interacts with DCAF1, leading to recruitment of the CUL4A-RBX1-DDB1- DCAF1/VPRBP complex to ubiquitinate proteins and limit error-prone repair during V(D)J recombination. {ECO:0000269\|PubMed:19396172, ECO:0000269\|PubMed:22157821, ECO:0000269\|PubMed:8521468, ECO:0000269\|PubMed:9094713, ECO:0000269\|PubMed:9184213, ECO:0000269\|PubMed:9228952}.
Subcellular location:		Nucleus {ECO:0000255\|PROSITE-ProRule:PRU00820, ECO:0000269\|PubMed:8284210}.
Tissue specificity:		Maturing lymphoid cells and central nervous system.
Domain:		The RING-type zinc finger mediates the E3 ubiquitin-protein ligase activity. {ECO:0000269\|PubMed:19396172}.
Domain:		The NBD (nonamer binding) DNA-binding domain mediates the specific binding to the nonamer RSS motif by forming a tightly interwoven homodimer that binds and synapses 2 nonamer elements, with each NBD making contact with both DNA molecules. Each RSS is composed of well-conserved heptamer (consensus 5'-CACAGTG-3') and nonamer (consensus 5'-ACAAAAACC-3') sequences separated by a spacer of either 12 bp or 23 bp. {ECO:0000255\|PROSITE-ProRule:PRU00820, ECO:0000269\|PubMed:19396172}.
Ptm:		Autoubiquitinated in the presence of CDC34/UBCH3. {ECO:0000269\|PubMed:14671314, ECO:0000269\|PubMed:19118899}.
Disruption phenotype:		Mice display a severe combined immunodeficiency phenotype. The have a small lymphoid organs that do not contain mature B and T-lymphocytes. The arrest of B- and T-cell differentiation occurs at an early stage and correlates with the inability to perform V(D)J recombination. The frequency of homologous immunoglobulin pairing is much lower. No obvious neuroanatomical or behavioral abnormalities have been observed. {ECO:0000269\|PubMed:1547488}.
Similarity:		Belongs to the RAG1 family. {ECO:0000255\|PROSITE- ProRule:PRU00820}.