1i3a: Difference between revisions

Revision as of 14:07, 21 February 2008

RNASE HII FROM ARCHAEOGLOBUS FULGIDUS WITH COBALT HEXAMMINE CHLORIDE

OverviewOverview

DNA replication and cellular survival requires efficient removal of RNA primers during lagging strand DNA synthesis. In eukaryotes, RNA primer removal is initiated by type 2 RNase H, which specifically cleaves the RNA portion of an RNA-DNA/DNA hybrid duplex. This conserved type 2 RNase H family of replicative enzymes shares little sequence similarity with the well-characterized prokaryotic type 1 RNase H enzymes, yet both possess similar enzymatic properties. Crystal structures and structure-based mutational analysis of RNase HII from Archaeoglobus fulgidus, both with and without a bound metal ion, identify the active site for type 2 RNase H enzymes that provides the general nuclease activity necessary for catalysis. The two-domain architecture of type 2 RNase H creates a positively charged binding groove and links the unique C-terminal helix-loop-helix cap domain to the active site catalytic domain. This architectural arrangement apparently couples directional A-form duplex binding, by a hydrogen-bonding Arg-Lys phosphate ruler motif, to substrate-discrimination, by a tyrosine finger motif, thereby providing substrate-specific catalytic activity. Combined kinetic and mutational analyses of structurally implicated substrate binding residues validate this binding mode. These structural and mutational results together suggest a molecular mechanism for type 2 RNase H enzymes for the specific recognition and cleavage of RNA in the RNA-DNA junction within hybrid duplexes, which reconciles the broad substrate binding affinity with the catalytic specificity observed in biochemical assays. In combination with a recent independent structural analysis, these results furthermore identify testable molecular hypotheses for the activity and function of the type 2 RNase H family of enzymes, including structural complementarity, substrate-mediated conformational changes and coordination with subsequent FEN-1 activity.

About this StructureAbout this Structure

1I3A is a Single protein structure of sequence from Archaeoglobus fulgidus with as ligand. Active as Ribonuclease H, with EC number 3.1.26.4 Full crystallographic information is available from OCA.

ReferenceReference

Structural biochemistry of a type 2 RNase H: RNA primer recognition and removal during DNA replication., Chapados BR, Chai Q, Hosfield DJ, Qiu J, Shen B, Tainer JA, J Mol Biol. 2001 Mar 23;307(2):541-56. PMID:11254381

Page seeded by OCA on Thu Feb 21 13:07:35 2008

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OCA

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-[[Image:1i3a.gif|left|200px]]<br /><applet load="1i3a" size="450" color="white" frame="true" align="right" spinBox="true"
+[[Image:1i3a.gif|left|200px]]<br /><applet load="1i3a" size="350" color="white" frame="true" align="right" spinBox="true"
 caption="1i3a, resolution 2.15&Aring;" />
 '''RNASE HII FROM ARCHAEOGLOBUS FULGIDUS WITH COBALT HEXAMMINE CHLORIDE'''<br />
 ==Overview==
-DNA replication and cellular survival requires efficient removal of RNA, primers during lagging strand DNA synthesis. In eukaryotes, RNA primer, removal is initiated by type 2 RNase H, which specifically cleaves the RNA, portion of an RNA-DNA/DNA hybrid duplex. This conserved type 2 RNase H, family of replicative enzymes shares little sequence similarity with the, well-characterized prokaryotic type 1 RNase H enzymes, yet both possess, similar enzymatic properties. Crystal structures and structure-based, mutational analysis of RNase HII from Archaeoglobus fulgidus, both with, and without a bound metal ion, identify the active site for type 2 RNase H, enzymes that provides the general nuclease activity necessary for, catalysis. The two-domain architecture of type 2 RNase H creates a, positively charged binding groove and links the unique C-terminal, helix-loop-helix cap domain to the active site catalytic domain. This, architectural arrangement apparently couples directional A-form duplex, binding, by a hydrogen-bonding Arg-Lys phosphate ruler motif, to, substrate-discrimination, by a tyrosine finger motif, thereby providing, substrate-specific catalytic activity. Combined kinetic and mutational, analyses of structurally implicated substrate binding residues validate, this binding mode. These structural and mutational results together, suggest a molecular mechanism for type 2 RNase H enzymes for the specific, recognition and cleavage of RNA in the RNA-DNA junction within hybrid, duplexes, which reconciles the broad substrate binding affinity with the, catalytic specificity observed in biochemical assays. In combination with, a recent independent structural analysis, these results furthermore, identify testable molecular hypotheses for the activity and function of, the type 2 RNase H family of enzymes, including structural, complementarity, substrate-mediated conformational changes and, coordination with subsequent FEN-1 activity.
+DNA replication and cellular survival requires efficient removal of RNA primers during lagging strand DNA synthesis. In eukaryotes, RNA primer removal is initiated by type 2 RNase H, which specifically cleaves the RNA portion of an RNA-DNA/DNA hybrid duplex. This conserved type 2 RNase H family of replicative enzymes shares little sequence similarity with the well-characterized prokaryotic type 1 RNase H enzymes, yet both possess similar enzymatic properties. Crystal structures and structure-based mutational analysis of RNase HII from Archaeoglobus fulgidus, both with and without a bound metal ion, identify the active site for type 2 RNase H enzymes that provides the general nuclease activity necessary for catalysis. The two-domain architecture of type 2 RNase H creates a positively charged binding groove and links the unique C-terminal helix-loop-helix cap domain to the active site catalytic domain. This architectural arrangement apparently couples directional A-form duplex binding, by a hydrogen-bonding Arg-Lys phosphate ruler motif, to substrate-discrimination, by a tyrosine finger motif, thereby providing substrate-specific catalytic activity. Combined kinetic and mutational analyses of structurally implicated substrate binding residues validate this binding mode. These structural and mutational results together suggest a molecular mechanism for type 2 RNase H enzymes for the specific recognition and cleavage of RNA in the RNA-DNA junction within hybrid duplexes, which reconciles the broad substrate binding affinity with the catalytic specificity observed in biochemical assays. In combination with a recent independent structural analysis, these results furthermore identify testable molecular hypotheses for the activity and function of the type 2 RNase H family of enzymes, including structural complementarity, substrate-mediated conformational changes and coordination with subsequent FEN-1 activity.
 ==About this Structure==
-I3A is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Archaeoglobus_fulgidus Archaeoglobus fulgidus] with NCO as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Ribonuclease_H Ribonuclease H], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.1.26.4 3.1.26.4] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1I3A OCA].
+I3A is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Archaeoglobus_fulgidus Archaeoglobus fulgidus] with <scene name='pdbligand=NCO:'>NCO</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Ribonuclease_H Ribonuclease H], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.1.26.4 3.1.26.4] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1I3A OCA].
 ==Reference==
@@ Line 15: / Line 15: @@
 [[Category: Single protein]]
 [[Category: Chai, Q.]]
-[[Category: Chapados, B.R.]]
+[[Category: Chapados, B R.]]
-[[Category: Hosfield, D.J.]]
+[[Category: Hosfield, D J.]]
 [[Category: Qiu, J.]]
 [[Category: Shen, B.]]
-[[Category: Tainer, J.A.]]
+[[Category: Tainer, J A.]]
 [[Category: NCO]]
 [[Category: helix-loop-helix]]
 [[Category: mixed beta sheet]]
-''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Tue Nov 20 17:01:33 2007''
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 13:07:35 2008''