Contributions of the C-terminal domain to poly(A)-specific ribonuclease (PARN) stability and self-association
Poly(A)-specific ribonuclease (PARN) catalyzes the degradation of mRNA poly(A) tail to regulate translation efficiency and mRNA decay in higher eukaryotic cells. The full-length PARN is a multi-domain protein containing the catalytic nuclease domain, the R3H domain, the RRM domain and the C-terminal...
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| Format: | Article |
| Language: | English |
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Elsevier
2019-07-01
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| Series: | Biochemistry and Biophysics Reports |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2405580818302954 |
| _version_ | 1818525493784215552 |
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| author | Guang-Jun He Yong-Bin Yan |
| author_facet | Guang-Jun He Yong-Bin Yan |
| author_sort | Guang-Jun He |
| collection | DOAJ |
| description | Poly(A)-specific ribonuclease (PARN) catalyzes the degradation of mRNA poly(A) tail to regulate translation efficiency and mRNA decay in higher eukaryotic cells. The full-length PARN is a multi-domain protein containing the catalytic nuclease domain, the R3H domain, the RRM domain and the C-terminal intrinsically unstructured domain (CTD). The roles of the three well-structured RNA-binding domains have been extensively studied, while little is known about CTD. In this research, the impact of CTD on PARN stability and aggregatory potency was studied by comparing the thermal inactivation and denaturation behaviors of full-length PARN with two N-terminal fragments lacking CTD. Our results showed that K+ induced additional regular secondary structures and enhanced PARN stability against heat-induced inactivation, unfolding and aggregation. CTD prevented PARN from thermal inactivation but promoted thermal aggregation to initiate at a temperature much lower than that required for inactivation and unfolding. Blue-shift of Trp fluorescence during thermal transitions suggested that heat treatment induced rearrangements of domain organizations. CTD amplified the stabilizing effect of K+, implying the roles of CTD was mainly achieved by electrostatic interactions. These results suggested that CTD might dynamically interact with the main body of the molecule and release of CTD promoted self-association via electrostatic interactions. Keywords: Deadenylase, Enzyme thermal stability, Enzyme thermal aggregation, Intrinsically disordered domain, Poly(A)-Specific ribonuclease (PARN), Self-association |
| first_indexed | 2024-12-11T06:09:55Z |
| format | Article |
| id | doaj.art-af85495add8d4e51b533ee7be71861f9 |
| institution | Directory Open Access Journal |
| issn | 2405-5808 |
| language | English |
| last_indexed | 2024-12-11T06:09:55Z |
| publishDate | 2019-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Biochemistry and Biophysics Reports |
| spelling | doaj.art-af85495add8d4e51b533ee7be71861f92022-12-22T01:18:09ZengElsevierBiochemistry and Biophysics Reports2405-58082019-07-0118Contributions of the C-terminal domain to poly(A)-specific ribonuclease (PARN) stability and self-associationGuang-Jun He0Yong-Bin Yan1State Key Laboratory of Membrane Biology, School of Life Sciences, Tsinghua University, Beijing, 100084, ChinaCorresponding author. School of Life Sciences, Tsinghua University, Beijing, 100084, China.; State Key Laboratory of Membrane Biology, School of Life Sciences, Tsinghua University, Beijing, 100084, ChinaPoly(A)-specific ribonuclease (PARN) catalyzes the degradation of mRNA poly(A) tail to regulate translation efficiency and mRNA decay in higher eukaryotic cells. The full-length PARN is a multi-domain protein containing the catalytic nuclease domain, the R3H domain, the RRM domain and the C-terminal intrinsically unstructured domain (CTD). The roles of the three well-structured RNA-binding domains have been extensively studied, while little is known about CTD. In this research, the impact of CTD on PARN stability and aggregatory potency was studied by comparing the thermal inactivation and denaturation behaviors of full-length PARN with two N-terminal fragments lacking CTD. Our results showed that K+ induced additional regular secondary structures and enhanced PARN stability against heat-induced inactivation, unfolding and aggregation. CTD prevented PARN from thermal inactivation but promoted thermal aggregation to initiate at a temperature much lower than that required for inactivation and unfolding. Blue-shift of Trp fluorescence during thermal transitions suggested that heat treatment induced rearrangements of domain organizations. CTD amplified the stabilizing effect of K+, implying the roles of CTD was mainly achieved by electrostatic interactions. These results suggested that CTD might dynamically interact with the main body of the molecule and release of CTD promoted self-association via electrostatic interactions. Keywords: Deadenylase, Enzyme thermal stability, Enzyme thermal aggregation, Intrinsically disordered domain, Poly(A)-Specific ribonuclease (PARN), Self-associationhttp://www.sciencedirect.com/science/article/pii/S2405580818302954 |
| spellingShingle | Guang-Jun He Yong-Bin Yan Contributions of the C-terminal domain to poly(A)-specific ribonuclease (PARN) stability and self-association Biochemistry and Biophysics Reports |
| title | Contributions of the C-terminal domain to poly(A)-specific ribonuclease (PARN) stability and self-association |
| title_full | Contributions of the C-terminal domain to poly(A)-specific ribonuclease (PARN) stability and self-association |
| title_fullStr | Contributions of the C-terminal domain to poly(A)-specific ribonuclease (PARN) stability and self-association |
| title_full_unstemmed | Contributions of the C-terminal domain to poly(A)-specific ribonuclease (PARN) stability and self-association |
| title_short | Contributions of the C-terminal domain to poly(A)-specific ribonuclease (PARN) stability and self-association |
| title_sort | contributions of the c terminal domain to poly a specific ribonuclease parn stability and self association |
| url | http://www.sciencedirect.com/science/article/pii/S2405580818302954 |
| work_keys_str_mv | AT guangjunhe contributionsofthecterminaldomaintopolyaspecificribonucleaseparnstabilityandselfassociation AT yongbinyan contributionsofthecterminaldomaintopolyaspecificribonucleaseparnstabilityandselfassociation |