| RRC ID |
5775
|
| Author |
Yoshinari S, Shiba T, Inaoka DK, Itoh T, Kurisu G, Harada S, Kita K, Watanabe Y.
|
| Title |
Functional importance of crenarchaea-specific extra-loop revealed by an X-ray structure of a heterotetrameric crenarchaeal splicing endonuclease.
|
| Journal |
Nucleic Acids Res
|
| Abstract |
Archaeal splicing endonucleases (EndAs) are currently classified into three groups. Two groups require a single subunit protein to form a homodimer or homotetramer. The third group requires two nonidentical protein components for the activity. To elucidate the molecular architecture of the two-subunit EndA system, we studied a crenarchaeal splicing endonuclease from Pyrobaculum aerophilum. In the present study, we solved a crystal structure of the enzyme at 1.7-A resolution. The enzyme adopts a heterotetrameric form composed of two catalytic and two structural subunits. By connecting the structural and the catalytic subunits of the heterotetrameric EndA, we could convert the enzyme to a homodimer that maintains the broad substrate specificity that is one of the characteristics of heterotetrameric EndA. Meanwhile, a deletion of six amino acids in a Crenarchaea-specific loop abolished the endonuclease activity even on a substrate with canonical BHB motif. These results indicate that the subunit architecture is not a major factor responsible for the difference of substrate specificity between single- and two-subunit EndA systems. Rather, the structural basis for the broad substrate specificity is built into the crenarchaeal splicing endonuclease itself.
|
| Volume |
37(14)
|
| Pages |
4787-98
|
| Published |
2009-8-1
|
| DOI |
10.1093/nar/gkp506
|
| PII |
gkp506
|
| PMID |
19515941
|
| PMC |
PMC2724299
|
| MeSH |
Amino Acid Sequence
Crystallography, X-Ray
Endoribonucleases / chemistry*
Endoribonucleases / genetics
Endoribonucleases / metabolism
Models, Molecular
Molecular Sequence Data
Protein Engineering
Protein Multimerization
Protein Subunits / chemistry
Pyrobaculum / enzymology*
Sequence Deletion
Substrate Specificity
|
| IF |
11.502
|
| Times Cited |
17
|
|
WOS Category
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| Resource |
| General Microbes |
JCM9630 |
