Posttranscriptional gene regulation and specific binding of the nonhistone protein HMG-I by the 3' untranslated region of bovine interleukin 2 cDNA.

AbstractPosttranscriptional gene regulation includes mRNA transport, localization, translation, and regulation of mRNA stability. CPEB (cytoplasmic polyadenylation element binding) family proteins bind to specific sites within the 3′-untranslated region and mediate poly- and deadenylation of transcripts, activating or repressing protein synthesis. As part of ribonucleoprotein complexes, the CPEB proteins participate in mRNA transport and localization to different sub-cellular compartments. The CPEB proteins are evolutionarily conserved and have similar functions in vertebrates and invertebrates. In the nervous system, the CPEB proteins are involved in cell division, neural development, learning, and memory. Here we consider the functional features of these proteins in the nervous system of phylogenetically distant organisms: Drosophila, a well-studied model, and mammals. Disruption of the CPEB proteins functioning is associated with various pathologies, such as autism spectrum disorder and brain cancer. At the same time, CPEB gene regulation can provide for a recovery of the brain function in patients with fragile X syndrome and Huntington's disease, making the CPEB genes promising targets for gene therapy.

Download Full-text

Specific binding of chloroplast proteins in vitro to the 3' untranslated region of spinach chloroplast petD mRNA

Molecular and Cellular Biology ◽

10.1128/mcb.11.9.4380-4388.1991 ◽

1991 ◽

Vol 11 (9) ◽

pp. 4380-4388

Author(s):

H C Chen ◽

D B Stern

Keyword(s):

Untranslated Region ◽

Specific Binding ◽

Rna Stability ◽

Repeat Sequence ◽

Hairpin Structure ◽

Sequence Motif ◽

Spinach Chloroplast ◽

Higher Plant ◽

Chloroplast Proteins

A detailed analysis of RNA-protein complex formation in the 3' untranslated region of spinach chloroplast petD mRNA has been carried out. Five chloroplast proteins that interact with petD RNA in this region, which contains an inverted repeat sequence capable of forming a hairpin structure, have been identified. A 33-kDa protein recognizes specifically the double-stranded stem of the hairpin structure; mutations that disrupt base pairing at the base of the stem reduce or eliminate protein binding. A 57-kDa protein recognizes specifically an AU-rich sequence motif that is highly conserved in petD genes of different higher plant species. The 57-kDa protein and possibly the 33-kDa protein form stable complexes with petD RNA in vitro and may interact with each other. In addition, their interaction with petD RNA is highly sensitive to heparin. The three other proteins, of 100, 32, and 28 kDa, display little sequence or structural binding specificity apart from their preference for uridine-rich sequences. They also interact with the 3' untranslated regions of other chloroplast RNAs such as those of psbA and rbcL. The functions of these proteins in the regulation of petD gene expression, including possible roles in transcription termination and RNA stability, are discussed.

Download Full-text

Mixed tailing by TENT4A and TENT4B shields mRNA from rapid deadenylation

Science ◽

10.1126/science.aam5794 ◽

2018 ◽

Vol 361 (6403) ◽

pp. 701-704 ◽

Cited By ~ 43

Author(s):

Jaechul Lim ◽

Dongwan Kim ◽

Young-suk Lee ◽

Minju Ha ◽

Mihye Lee ◽

...

Keyword(s):

Gene Regulation ◽

Half Life ◽

Messenger Rna ◽

Mrna Translation ◽

Posttranscriptional Gene Regulation ◽

And Function ◽

Mrna Half Life ◽

In Cells

RNA tails play integral roles in the regulation of messenger RNA (mRNA) translation and decay. Guanylation of the poly(A) tail was discovered recently, yet the enzymology and function remain obscure. Here we identify TENT4A (PAPD7) and TENT4B (PAPD5) as the enzymes responsible for mRNA guanylation. Purified TENT4 proteins generate a mixed poly(A) tail with intermittent non-adenosine residues, the most common of which is guanosine. A single guanosine residue is sufficient to impede the deadenylase CCR4-NOT complex, which trims the tail and exposes guanosine at the 3′ end. Consistently, depletion of TENT4A and TENT4B leads to a decrease in mRNA half-life and abundance in cells. Thus, TENT4A and TENT4B produce a mixed tail that shields mRNA from rapid deadenylation. Our study unveils the role of mixed tailing and expands the complexity of posttranscriptional gene regulation.

Download Full-text

Eukaryotic Elongation Factor 1A Interacts with the Upstream Pseudoknot Domain in the 3′ Untranslated Region of Tobacco Mosaic Virus RNA

Journal of Virology ◽

10.1128/jvi.76.11.5678-5691.2002 ◽

2002 ◽

Vol 76 (11) ◽

pp. 5678-5691 ◽

Cited By ~ 75

Author(s):

Vladimir V. Zeenko ◽

Lyubov A. Ryabova ◽

Alexander S. Spirin ◽

Helen M. Rothnie ◽

Daniel Hess ◽

...

Keyword(s):

Tobacco Mosaic Virus ◽

Mosaic Virus ◽

Untranslated Region ◽

Mutational Analysis ◽

Specific Binding ◽

Elongation Factor ◽

Virus Rna ◽

Eukaryotic Elongation Factor ◽

Sequence Elements ◽

Positive Strand Rna

ABSTRACT The genomic RNA of tobacco mosaic virus (TMV), like that of other positive-strand RNA viruses, acts as a template for both translation and replication. The highly structured 3′ untranslated region (UTR) of TMV RNAs plays an important role in both processes; it is not polyadenylated but ends with a tRNA-like structure (TLS) preceded by a conserved upstream pseudoknot domain (UPD). The TLS of tobamoviral RNAs can be specifically aminoacylated and, in this state, can interact with eukaryotic elongation factor 1A (eEF1A)/GTP with high affinity. Using a UV cross-linking assay, we detected another specific binding site for eEF1A/GTP, within the UPDs of TMV and crucifer-infecting tobamovirus (crTMV), that does not require aminoacylation. A mutational analysis revealed that UPD pseudoknot conformation and some conserved primary sequence elements are required for this interaction. Its possible role in the regulation of tobamovirus gene expression and replication is discussed.

Download Full-text

Transcriptional and Posttranscriptional Regulation of Insulin-Like Growth Factor Binding Protein-3 by Cyclic Adenosine 3′,5′-Monophosphate: Messenger RNA Stabilization Is Accompanied by Decreased Binding of a 42-kDa Protein to a Uridine-Rich Domain in the 3′-Untranslated Region

Molecular Endocrinology ◽

10.1210/mend.13.3.0252 ◽

1999 ◽

Vol 13 (3) ◽

pp. 495-504

Author(s):

N. E. Erondu ◽

J. Nwankwo ◽

Y. Zhong ◽

M. Boes ◽

B. Dake ◽

...

Keyword(s):

Growth Factor ◽

Untranslated Region ◽

Specific Binding ◽

Binding Protein ◽

Binding Activity ◽

Insulin Like Growth Factor ◽

Factor Binding ◽

Mdbk Cells ◽

Camp Induction ◽

Igfbp 3

Abstract The Madin Darby bovine kidney (MDBK) cell line was used to investigate the mechanisms underlying the cAMP regulation of insulin-like growth factor binding protein-3 (IGFBP-3) gene expression. Treatment of confluent monolayers either with forskolin or cAMP produced a 60- to 75-fold induction of IGFBP-3 mRNA and protein levels. This effect did not require new protein synthesis as inhibition of translation by cycloheximide actually caused a 2-fold increase in the cAMP induction. The rates of IGFBP-3 gene transcription, assessed by nuclear run-on assays, increased approximately 15-fold in cells exposed to cAMP. In addition, the half-life of the IGFBP-3 mRNA transcript was increased ∼3-fold in the presence of cAMP. Gel mobility shift and competition experiments revealed the specific binding of an approximately 42-kDa cytoplasmic protein factor to the 3′-untranslated region (3′-UTR) of the IGFBP-3 mRNA. A 21-nucleotide uridine-rich segment that contained no AUUUA motif was sufficient for the specific binding. The binding activity of this protein was reduced after cAMP treatment but was increased by phosphatase treatment. In conclusion, the cAMP induction of IGFBP-3 mRNA in MDBK cells occurred at both the transcriptional and posttranscriptional levels. The IGFBP-3 mRNA stabilization in MDBK cells probably involved the phosphorylation of a member of the family of U-rich region mRNA-binding proteins and is the first reported member whose RNA-binding activity is reduced by cAMP.

Download Full-text