Phosphorylation is required for high-affinity binding of DBP, a yeast mitochondrial site-specific RNA binding protein

2000 ◽  
Vol 37 (6) ◽  
pp. 356-363 ◽  
Author(s):  
Huilin Li ◽  
H. Peter Zassenhaus
Keyword(s):  
Rna Binding ◽  
Binding Protein ◽  
Rna Binding Protein ◽  
Affinity Binding ◽  
Site Specific ◽  
High Affinity Binding ◽  
High Affinity

scholarly journals Hepatitis C Virus Nonstructural Protein 5A (NS5A) Is an RNA-binding Protein

2005 ◽  
Vol 280 (43) ◽  
pp. 36417-36428 ◽  
Author(s):  
Luyun Huang ◽  
Jungwook Hwang ◽  
Suresh D. Sharma ◽  
Michele R. S. Hargittai ◽  
Yingfeng Chen ◽  
...  
Keyword(s):  
Hepatitis C ◽  
Rna Binding ◽  
Nonstructural Protein ◽  
Rna Binding Protein ◽  
Affinity Binding ◽  
Genome Replication ◽  
Subgenomic Replicon ◽  
High Affinity Binding ◽  
High Affinity ◽  
Nonstructural Protein 5A

Hepatitis C virus (HCV) nonstructural protein 5A (NS5A) has been shown to antagonize numerous cellular pathways, including the antiviral interferon-α response. However, the capacity of this protein to interact with the viral polymerase suggests a more direct role for NS5A in genome replication. In this study, we employed two bacterially expressed, soluble derivatives of NS5A to probe for novel functions of this protein. We find that NS5A has the capacity to bind to the 3′-ends of HCV plus and minus strand RNAs. The high affinity binding site for NS5A in the 3′-end of plus strand RNA maps to the polypyrimidine tract, an element known to be essential for genome replication and infectivity. NS5A has a preference for single-stranded RNA containing stretches of uridine or guanosine. Values for the equilibrium dissociation constants for high affinity binding sites were in the 10 nm range. Two-dimensional gel electrophoresis followed by Western blotting revealed the presence of unphosphorylated NS5A in Huh-7 cells stably expressing the subgenomic replicon. Moreover, RNA immunoprecipitation and NS5A pull-down experiments showed the capacity of replicon-derived NS5A to bind to synthetic RNA and the HCV genome, respectively. Deletion of all of the casein kinase II phosphorylation sites in NS5A supported stable replication of a subgenomic replicon in Huh-7. However, this derivative could not be labeled with inorganic phosphate, suggesting that extensive phosphorylation of NS5A is not required for the replication functions of NS5A. The discovery that NS5A is an RNA-binding protein defines a new functional target for development of agents to treat HCV infection and a new structural class of RNA-binding proteins.

scholarly journals Agrin Is a High-affinity Binding Protein of Dystroglycan in Non-muscle Tissue

1998 ◽  
Vol 273 (1) ◽  
pp. 600-605 ◽  
Author(s):  
Matthias Gesemann ◽  
Andrea Brancaccio ◽  
Beat Schumacher ◽  
Markus A. Ruegg
Keyword(s):  
Muscle Tissue ◽  
Binding Protein ◽  
Affinity Binding ◽  
High Affinity Binding ◽  
High Affinity

scholarly journals The human RNA-binding protein and E3 ligase MEX-3C binds the MEX-3–recognition element (MRE) motif with high affinity

2017 ◽  
Vol 292 (39) ◽  
pp. 16221-16234 ◽  
Author(s):  
Lingna Yang ◽  
Chongyuan Wang ◽  
Fudong Li ◽  
Jiahai Zhang ◽  
Anam Nayab ◽  
...  
Keyword(s):  
Rna Binding ◽  
Binding Protein ◽  
Rna Binding Protein ◽  
E3 Ligase ◽  
Recognition Element ◽  
High Affinity

scholarly journals N-Acylethanolamine Signaling in Tobacco Is Mediated by a Membrane-Associated, High-Affinity Binding Protein

2003 ◽  
Vol 131 (4) ◽  
pp. 1781-1791 ◽  
Author(s):  
Swati Tripathy ◽  
Kathryn Kleppinger-Sparace ◽  
Richard A. Dixon ◽  
Kent D. Chapman
Keyword(s):  
Binding Protein ◽  
Affinity Binding ◽  
High Affinity Binding ◽  
High Affinity

scholarly journals The neuronal RNA binding protein Nova-1 recognizes specific RNA targets in vitro and in vivo.

1997 ◽  
Vol 17 (6) ◽  
pp. 3194-3201 ◽  
Author(s):  
R J Buckanovich ◽  
R B Darnell
Keyword(s):  
Rna Binding ◽  
Binding Protein ◽  
Rna Binding Protein ◽  
Motor Dysfunction ◽  
High Affinity ◽  
Loop Region ◽  
Nuclear Rna ◽  
Rna Ligands

Nova-1, an autoantigen in paraneoplastic opsoclonus myoclonus ataxia (POMA), a disorder associated with breast cancer and motor dysfunction, is a neuron-specific nuclear RNA binding protein. We have identified in vivo Nova-1 RNA ligands by combining affinity-elution-based RNA selection with protein-RNA immunoprecipitation. Starting with a pool of approximately 10(15) random 52-mer RNAs, we identified long stem-loop RNA ligands that bind to Nova-1 with high affinity (Kd of approximately 2 nM). The loop region of these RNAs harbors a approximately 15-bp pyrimidine-rich element [UCAU(N)(0-2)]3 which is essential for Nova-1 binding. Mutagenesis studies defined the third KH domain of Nova-1 and the [UCAU(N)(0-2)]3 element as necessary for in vitro binding. Consensus [UCAU (N)(0-2)], elements were identified in two neuronal pre-mRNAs, one encoding the inhibitory glycine receptor alpha2 (GlyR alpha2) and a second encoding Nova-1 itself. Nova-1 protein binds these RNAs with high affinity and specificity in vitro, and this binding can be blocked by POMA antisera. Moreover, both Nova-1 and GlyR alpha2 pre-mRNAs specifically coimmunoprecipitated with Nova-1 protein from brain extracts. Thus, Nova-1 functions as a sequence-specific nuclear RNA binding protein in vivo; disruption of the specific interaction between Nova-1 and GlyR alpha2 pre-mRNA may underlie the motor dysfunction seen in POMA.

scholarly journals Efficient homing of antibody-secreting cells to the bone marrow requires RNA-binding protein ZFP36L1

2020 ◽  
Vol 218 (3) ◽  
Author(s):  
Alexander Saveliev ◽  
Sarah E. Bell ◽  
Martin Turner
Keyword(s):  
Bone Marrow ◽  
Rna Binding ◽  
Binding Protein ◽  
Rna Binding Protein ◽  
Receptor Kinase ◽  
High Affinity ◽  
Guidance Receptors ◽  
G Protein Coupled ◽  
Late Stages ◽  
Antibody Secreting Cells

Cell migration relies on coordinated activity of chemotactic and guidance receptors. Here, we report a specific role for the RNA-binding protein ZFP36L1 in limiting the abundance of molecules involved in the homing of antibody-secreting cells (ASCs) to the bone marrow (BM). In the absence of ZFP36L1, ASCs build up in the spleen and the liver and show diminished accumulation in the BM. ZFP36L1 facilitates migration by directly regulating G protein–coupled receptor kinase 2 (GRK2) and the integrin chains α4 and β1 in splenic ASCs. Expression of CXCR4 and of the integrins α4 and β1 is differentially regulated on ASCs produced at the early and late stages of the immune response. Consequently, deletion of the Zfp36l1 gene has a stronger effect on BM accumulation of high-affinity ASCs formed late in the response. Thus, ZFP36L1 is an integral part of the regulatory network controlling gene expression during ASC homing.

RAT MYOMETRIAL ACTIVITY IN VIVO: EFFECTS OF OESTRADIOL-17β AND PROGESTERONE IN RELATION TO THE CONCENTRATIONS OF CYTOPLASMIC PROGESTERONE RECEPTORS

1978 ◽  
Vol 78 (1) ◽  
pp. 103-117 ◽  
Author(s):  
SANDRA J. DOWNING ◽  
S. J. LYE ◽  
JANE M. C. BRADSHAW ◽  
D. G. PORTER
Keyword(s):  
Binding Protein ◽  
Progesterone Receptors ◽  
Affinity Binding ◽  
Pregnant Rats ◽  
Pregnant Rat ◽  
High Affinity Binding ◽  
High Affinity ◽  
Repeated Doses ◽  
In Vivo Effects

The amplitude, frequency and rate of rise of intra-uterine pressure cycles in rats (postpartum, ovariectomized) were unaffected by treatment with progesterone. Amplitude was also unaffected by a combination of treatments with progesterone and oestradiol-17β, which was adequate to ensure the survival of 84% of foetuses in ovariectomized pregnant rats. The failure of progesterone to influence myometrial activity could not be attributed to a lack of 'true' progesterone receptors since these were present in the myometria of the test animals in concentrations exceeding those of oestrous animals. Evidence was obtained which suggested that a high-affinity binding protein, different from the 'true' receptor may predominate in the myometrium of the pregnant rat. Oestradiol-17β in single or repeated doses of from 0·25 to 5 μg, however, was found to reduce the frequency of pressure cycles but to increase significantly their rate of rise of pressure. There was a latency of 6–8 h in these effects of oestradiol. The possibility that inhibition of the myometrium by oestrogen may play a part in the preparation for parturition is discussed.

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