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Literature summary for 3.1.26.5 extracted from

  • Hands-Taylor, K.L.; Martino, L.; Tata, R.; Babon, J.J.; Bui, T.T.; Drake, A.F.; Beavil, R.L.; Pruijn, G.J.; Brown, P.R.; Conte, M.R.
    Heterodimerization of the human RNase P/MRP subunits Rpp20 and Rpp25 is a prerequisite for interaction with the P3 arm of RNase MRP RNA (2010), Nucleic Acids Res., 38, 4052-4066.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
cDNAs of full-length Rpp20 and Rpp25 cloned into pCR4-TOPO. Full-length Rpp25 (encompassing residues 1-199) subcloned with an N-terminal hexahistidine tag into a pPROEX-HTb expression vector using Nco1/Not1 restriction sites. Full-length Rpp20 (residues 1-140) subcloned into a pET-30 expression vector. The pET-30 vector modified to bear a TEV cleavage site to remove the N-terminal hexahistidine tag. Mutant Rpp20(35-140) amplified from pET-30 Rpp20 to introduce 5' Asc1 and 3' Not1 sites and then ligated into a modified version of pET-15b vector with an Asc1 site inserted directly after the his tag. Fragments Rpp20, mutant Rpp20(16-140), mutant Rpp25(25-170) and co-expressed Rpp20/Rpp25 (in which Rpp20 bears the his tag) subcloned in pETDuet-1 vector with an N-terminal hexahistidine tag. Mutant Rpp25(25-170) also subcloned in a pCDFDuet-1 vector also with an N-terminal hexahistidine tag. Rpp25, Rpp20 (from pET-30 vector) and mutant Rpp20(35-140) expressed in Escherichia coli strain BL21(DE3)pLysS. All the other proteins subcloned into pETDuet-1 or pCDFDuet-1 vectors expressed in Escherichia coli KRX strain Homo sapiens

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
13200
-
2 * 16000, full-length Rpp20, gel filtration. 2 * 14100, Rpp20(16-140), gel filtration. 2 * 13200, HisRpp20(35-140), gel filtration Homo sapiens
14100
-
2 * 16000, full-length Rpp20, gel filtration. 2 * 14100, Rpp20(16-140), gel filtration. 2 * 13200, HisRpp20(35-140), gel filtration Homo sapiens
15400
-
mutant Rpp25(25-170), gel filtration Homo sapiens
16000
-
2 * 16000, full-length Rpp20, gel filtration. 2 * 14100, Rpp20(16-140), gel filtration. 2 * 13200, HisRpp20(35-140), gel filtration Homo sapiens
20600
-
mutant Rpp25, gel filtration Homo sapiens
20700
-
mutant Rpp25(25-170), sequence analysis Homo sapiens
26400
-
mutant HisRpp20(35-140), gel filtration Homo sapiens
28100
-
mutant Rpp20(16-140), gel filtration Homo sapiens
28600
-
mutant Rpp20(35-140)/Rpp25(25-170), gel filtration Homo sapiens
31100
-
mutant Rpp20(35-140)/Rpp25(25-170), sequence analysis Homo sapiens
31500
-
mutant Rpp20/Rpp25(25-170), gel filtration Homo sapiens
32100
-
full-length Rpp20, gel filtration Homo sapiens
36600
-
mutant Rpp20/Rpp25, gel filtration Homo sapiens
39200
-
mutant Rpp25, sequence analysis Homo sapiens
39900
-
mutant HisRpp20(35-140), sequence analysis Homo sapiens
41100
-
mutant Rpp20/Rpp25(25-170), sequence analysis Homo sapiens
44400
-
mutant [Rpp20(35-140)-Rpp25(25-170)]/P3 RNA, gel filtration Homo sapiens
45300
-
mutant Rpp20(16-140), sequence analysis Homo sapiens
47000
-
full-length Rpp20, sequence analysis Homo sapiens
52500
-
mutant [Rpp20-Rpp25]/P3 RNA, gel filtration Homo sapiens
53800
-
mutant Rpp20/Rpp25, sequence analysis Homo sapiens
56400
-
mutant [Rpp20(35-140)-Rpp25(25-170)]/P3 RNA, sequence analysis Homo sapiens
71400
-
mutant [Rpp20-Rpp25]/P3 RNA, sequence analysis Homo sapiens

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-

Source Tissue

Source Tissue Comment Organism Textmining

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information Rpp20 and Rpp25 interact with the P3 arm of RNase MRP RNA in a highly synergic fashion. Rpp20 and Rpp25 interact with the P3 RNA as a heterodimer, which is formed prior to RNA binding Homo sapiens ?
-
?

Subunits

Subunits Comment Organism
heterodimer Rpp20 and Rpp25, ITC-200 microcalorimeter experiments Homo sapiens
homodimer 2 * 16000, full-length Rpp20, gel filtration. 2 * 14100, Rpp20(16-140), gel filtration. 2 * 13200, HisRpp20(35-140), gel filtration Homo sapiens
monomer Rpp25 in solution Homo sapiens

Synonyms

Synonyms Comment Organism
RNase P
-
Homo sapiens
RNase P/MRP
-
Homo sapiens
Rpp20
-
Homo sapiens
Rpp25
-
Homo sapiens

General Information

General Information Comment Organism
malfunction in both deletion mutants Rpp20(16-140) and Rpp20(35-140) the global thermodynamics of the interaction with Rpp25 is seemingly unaffected. Thermodynamic signature of the association is also fully preserved between the Rpp25(25-170) mutant and all available versions of Rpp20. Regions within the mutants Rpp20(35-140) and Rpp25(25-170) are sufficient for mutual interaction, thus this recognition can be mediated largely, if not exclusively, by the Alba-type core domains Homo sapiens
physiological function strong interaction between Rpp25 and Rpp20. Rpp20 and Rpp25 interact with the P3 arm of RNase MRP RNA in a highly synergic fashion. Rpp20 and Rpp25 interact with the P3 RNA as a heterodimer, which is formed prior to RNA binding. Association between Rpp20 and Rpp25 has no detectable influence on their secondary/tertiary structure. The association reaction results in a large loss of solvent-accessible area. N- and C-terminal regions of Rpp25 and the N-terminal tail of Rpp20 are not involved in mutual recognition Homo sapiens