Messenger RNA population analysis during erythroid differentiation: A kinetical approach

Bastos, Roberto N.; Volloch, Zeev; Aviv, Haim

doi:10.1016/s0022-2836(77)80068-5

Cited by 86 publications

(57 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Although the high stability of human ␤-globin mRNA has been repeatedly demonstrated (2,4,36,42,62,63) and the importance of this characteristic to normal ␤-globin expression firmly established (10,64), the relevant molecular mechanisms have yet to be defined. The current article provides original evidence that this process is likely to require cis determinants and trans-acting factors that have not previously been appreciated.…”

Section: Discussionmentioning

confidence: 99%

“…In vivo analyses estimate a half-life for human ␣-globin mRNA of between 24 and 60 h (47,62,63,74), while similar studies with cultured NIH 3T3 and murine erythroleukemia (MEL) cells (2,36,42), primary mouse hematopoietic cells (4), and human erythroid progenitors (62,63) suggest a half-life value for human ␤-globin mRNA that exceeds 16 to 20 h. Globin mRNAs survive, and continue to translate at high levels, for as long as a week following nuclear condensation and extrusion in transcriptionally silent erythroid progenitor cells. As might be anticipated, mutations that impair the normal stabilities of globin mRNAs can severely impact the levels of their encoded proteins.…”

mentioning

confidence: 89%

See 1 more Smart Citation

A Nucleolin-Binding 3′ Untranslated Region Element Stabilizes β-Globin mRNA In Vivo

Jiang¹,

Xu²,

Russell

2006

Molecular and Cellular Biology

107

View full text Add to dashboard Cite

The normal expression of human ␤ globin is critically dependent upon the constitutively high stability of its encoding mRNA. Unlike with ␣-globin mRNA, the specific cis-acting determinants and trans-acting factors that participate in stabilizing ␤-globin mRNA are poorly described. The current work uses a linker-scanning strategy to identify a previously unknown determinant of mRNA stability within the ␤-globin 3 untranslated region (3UTR). The new determinant is positioned on an mRNA half-stem opposite a pyrimidine-rich sequence targeted by ␣CP/hnRNP-E, a factor that plays a critical role in stabilizing human ␣-globin mRNA. Mutations within the new determinant destabilize ␤-globin mRNA in intact cells while also ablating its 3UTR-specific interaction with the polyfunctional RNA-binding factor nucleolin. We speculate that 3UTR-bound nucleolin enhances mRNA stability by optimizing ␣CP access to its functional binding site. This model is favored by in vitro evidence that ␣CP binding is enhanced both by cis-acting stem-destabilizing mutations and by the trans-acting effects of supplemental nucleolin. These studies suggest a mechanism for ␤-globin mRNA stability that is related to, but distinct from, the mechanism that stabilizes human ␣-globin mRNA.Erythroid cells accumulate hemoglobin through a process that is critically dependent upon the high stabilities of mRNAs that encode their constituent ␣-and ␤-globin subunits (10, 64). In vivo analyses estimate a half-life for human ␣-globin mRNA of between 24 and 60 h (47,62,63,74), while similar studies with cultured NIH 3T3 and murine erythroleukemia (MEL) cells (2, 36, 42), primary mouse hematopoietic cells (4), and human erythroid progenitors (62, 63) suggest a half-life value for human ␤-globin mRNA that exceeds 16 to 20 h. Globin mRNAs survive, and continue to translate at high levels, for as long as a week following nuclear condensation and extrusion in transcriptionally silent erythroid progenitor cells. As might be anticipated, mutations that impair the normal stabilities of globin mRNAs can severely impact the levels of their encoded proteins. For example, an mRNA-destabilizing mutation reduces the expression of ␣ Constant Spring to less than 2% of normal levels (45,51,80), resulting in a clinically important form of thalassemia characterized by a substantial imbalance in ␣-and ␤-globin chain accumulation (10, 64).The cis-acting determinants and trans-acting factors that participate in regulating ␣-globin mRNA stability have recently been identified, and the relevant molecular mechanisms have been described in detail. Mutational analyses carried out with cultured cells (80, 81) and with animal models (52, 66) clearly demonstrate the importance of the 3Ј untranslated region (3ЈUTR) to the constitutively high stability of ␣-globin mRNA (45). Other studies have mapped this characteristic to a phylogenically conserved, 16-nucleotide (nt) C/U-rich element in this region (39,75,76). The cis-acting pyrimidine-rich element (PRE) assembles an mRNP "␣-complex" that compri...

show abstract

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 89%

A Nucleolin-Binding 3′ Untranslated Region Element Stabilizes β-Globin mRNA In Vivo

Jiang¹,

Xu²,

Russell

2006

Molecular and Cellular Biology

107

View full text Add to dashboard Cite

show abstract

“…Globin genes are expressed in highly specialized, terminally differentiated erythrocytes (RBC) and can account for as much as 95% of the soluble proteins (17,43). During RBC differentiation the nucleus is extruded (17) and globin mRNAs increase from less than 1% to more than 98% of the total mRNA (6). This high-level accumulation of globin mRNA is dependent not only on increased transcription but also on the long half-lives of globin mRNAs, 24 to 60 h, and the degradation of nonglobin mRNAs (3,5,30,34,49,50).…”

mentioning

confidence: 99%

Erythroid cell-specific determinants of alpha-globin mRNA stability.

1994

View full text Add to dashboard Cite

Although globin mRNAs are considered prototypes of highly stable messages, the mechanisms responsible for their longevity remain largely undefined. As an initial step in identifying potential cis-acting elements or structures which contribute to their stability, we analyzed the defect in expression of a naturally occurring a2-globin mutant, aConstant Spring (CS). The CS mutation is a single-base change in the translation termination codon (I!AA-WCAA) that allows the ribosome to read through into the 3' nontranslated region (NTR). The presence of CS mRNA in transcriptionally active erythroid precursors and its absence (relative to normal a-globin mRNA) in the more differentiated transcriptionally silent erythrocytes suggest that this mutation disrupts some feature of the a-globin mRNA required for its stability. Using a transient transfection system, we demonstrate that in murine erythroleukemia cells the CS mRNA is unstable compared with the normal a2-globin mRNA. The analyses of several other naturally occurring and site-directed mutant a-globin genes in murine erythroleukemia cells indicate that entry of a translating ribosome into the 3' NTR targets the message for accelerated degradation in erythroid cells. In contrast, both the CS and a2-globin mRNAs are stable in several nonerythroid cell lines. These results suggest that translational readthrough disrupts a determinant associated with the a2-globin 3' NTR which is required for mRNA stability in erythroid cells.While the major roles of transcription and processing in the control of gene expression have long been acknowledged (13), a number of systems which demonstrate a primary role for regulation at the level of mRNA stability have recently been described (reviewed in references 2, 7, 42, and 44). In eucaryotes, mRNAs have a wide range of half-lives, from as short as a few minutes to as long as several days. These differences in mRNA stability can have significant impacts on the levels of gene expression (20,48). A variety of elements and mechanisms appear to be responsible for modulating mRNA halflives. Features governing the turnover of individual mRNAs have been attributed to specific cis-acting RNA sequences and structures and to trans-acting proteins (10,27,36,41,45,46,55,57,63). An additional level of complexity is possible as an mRNA is translated, since the interaction of cis-and transacting elements with the ribosome and its associated components or with higher-order RNA structures could modulate mRNA turnover (47,52).The at-and P-globin gene clusters are useful model systems to study mechanisms responsible for mRNA stability. Globin genes are expressed in highly specialized, terminally differentiated erythrocytes (RBC) and can account for as much as 95% of the soluble proteins (17,43). During RBC differentiation the nucleus is extruded (17) and globin mRNAs increase from less than 1% to more than 98% of the total mRNA (6). This high-level accumulation of globin mRNA is dependent not only on increased transcription but also on the long half-lives...

show abstract

“…In spite of difficulties in efficiently chasing radioactively labeled uridine from the UTP pool in some cell types (66; unpublished observations), simply washing and suspending cells in medium containing high concentrations of uridine and cytidine sometimes achieves an effective dilution (5,36). When S49 cells labeled for 30 min with 500 pLCi of [3H]uridine per ml were washed and suspended in medium containing 10 mM uridine and cytidine, the incorporation of [3H]uridine into acid-soluble molecules ceased after the washes.…”

mentioning

confidence: 99%

Inhibition of protein synthesis stabilizes histone mRNA.

Stimac¹,

Groppi²,

Coffino³

1984

Mol. Cell. Biol.

118

View full text Add to dashboard Cite

The inhibition of protein synthesis in exponentially growing S49 cells leads to a specific fivefold increase in histone mRNA in 30 min. The rate of transcription of histone mRNA, measured in intact or digitoninpermeabilized cells, is increased slightly, if at all, by cycloheximide inhibition of protein synthesis. Both approach-to-equilibrium labeling and pulse-chase experiments show that cycloheximide prolongs histone mRNA half-life from approximately 30 min to >2 h. Histone mRNA made before the addition of cycloheximide becomes stable after the inhibition of protein synthesis, whereas removal of the inhibitor is followed by rapid degradation of histone mRNA. This suggests that the increased stability of histone mRNA during inhibition of protein synthesis results not from alteration of the structure of the mRNA, but from the loss of an activity in the cell which regulates histone mRNA turnover.

show abstract

Messenger RNA population analysis during erythroid differentiation: A kinetical approach

Cited by 86 publications

References 32 publications

A Nucleolin-Binding 3′ Untranslated Region Element Stabilizes β-Globin mRNA In Vivo

A Nucleolin-Binding 3′ Untranslated Region Element Stabilizes β-Globin mRNA In Vivo

Erythroid cell-specific determinants of alpha-globin mRNA stability.

Inhibition of protein synthesis stabilizes histone mRNA.

Contact Info

Product

Resources

About