Brain-specific polyA- transcripts are detected in polyA+ RNA: do complex polyA- brain RNAs really exist?

Fung, BP; Brilliant, MH; Chikaraishi, DM

doi:10.1523/jneurosci.11-03-00701.1991

Cited by 12 publications

(10 citation statements)

References 29 publications

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“…The RNase protections were performed as described by Fung et al (1991Fung et al ( , 1992 with some minor modifications. The RNase protection probe was constructed as follows.…”

Section: Methodsmentioning

confidence: 99%

The Cyclic AMP Response Element Directs Tyrosine Hydroxylase Expression in Catecholaminergic Central and Peripheral Nervous System Cell Lines from Transgenic Mice

Lazaroff

Patankar

Yoon

et al. 1995

Journal of Biological Chemistry

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Enhancer elements regulating the neuronal gene, tyrosine hydroxylase (TH), were identified in TH-expressing peripheral nervous system PATH and central nervous system CATH cell lines. Mutational analysis in which rat TH 5-flanking sequences directed chloramphenicol acetyltransferase (CAT) reporter gene expression demonstrated that mutating the cyclic AMP response element (CRE) at ؊45 base pair reduced expression by 80 -90%. A CRE linked to an enhancerless TH promoter fully supported expression. Cotransfection of a dominant-negative CREB protein reduced expression 50 -60%, suggesting that the CRE is bound by CREB or a CREB dimerization partner. Although mutating the AP1/dyad (AD) element at ؊205 base pair only modestly reduced CAT levels, AD minimal enhancer constructs gave 45-80% of wild type expression when positioned at ؊91 or ؊95. However, in its native context at ؊205, the AD could not support expression. In contrast, a CRE, moved from its normal position at ؊45 to ؊206, gave full activity. These results indicate that the CRE is critical for TH transcription in central nervous system CATH and peripheral nervous system PATH cells, whereas the AD is less important and its enhancer activity is contextand/or position-dependent. These results represent the first attempts to map regulatory elements directing TH expression in central nervous system cell lines. Tyrosine hydroxylase (TH)1 converts L-tyrosine to 3,4-dihydroxy-L-phenylalanine and is the first and rate-limiting enzyme in catecholamine synthesis (Nagatsu et al., 1964;Levitt et al., 1965). TH is expressed in specific cell types in the peripheral and central nervous systems. Sympathetic ganglia and chromaffin cells of the adrenal medulla are major sites of peripheral TH expression. In the central nervous system, TH-expressing neurons are located in the diencephalon, midbrain, brainstem, olfactory bulb, and retina (Bjorklund and Lindvall, 1984).TH activity is regulated at the protein level and the RNA level. Activation at the protein level is short term with a time course of less than 1 h and mainly occurs via phosphorylation of preexisting protein molecules, which increases TH activity (reviewed by Zigmond et al. (1989)). Induction of TH at the mRNA level is long term, resulting in increased mRNA levels for hours to days. Long term increases of TH activity are induced by various physiological stimuli including cyclic AMP (cAMP), epidermal growth factor, glucocorticoids, nerve growth factor, transsynaptic neuronal activity, and depolarization. These inducers have been shown to increase TH mRNA levels, and several studies indicate that they increase mRNA levels by inducing transcription: cAMP (Lewis et al

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“…The RNase protections were performed as described by Fung et al (1991Fung et al ( , 1992 with some minor modifications. The RNase protection probe was constructed as follows.…”

Section: Methodsmentioning

confidence: 99%

The Cyclic AMP Response Element Directs Tyrosine Hydroxylase Expression in Catecholaminergic Central and Peripheral Nervous System Cell Lines from Transgenic Mice

Lazaroff

Patankar

Yoon

et al. 1995

Journal of Biological Chemistry

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“…In general, nonpolyadenylated brain mRNA is probably biologically significant (Kaufmann et al, 1977;Chikaraishi et al, 1978;Morrison et al, 1979;Chikaraishi, 1979;Hahn et al, 1983). Some poly(A) Ϫ RNA may derive from poly(A) ϩ RNA by breakage during isolation (Fung et al, 1991); however, it is unlikely that Girk2D derives by random breakage since that would be expected to result in a smear on Northern blots. Also, the CNS specificity of Girk2D indicates that it probably is not generated by a random process.…”

Section: Fig 6 Expression In Testismentioning

confidence: 99%

Characterization of MurineGirk2Transcript Isoforms: Structure and Differential Expression

et al. 1998

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“…Nuclease protection experiments were performed as described (Fung et al, 1991). The nuclease protection probe was constructed as follows: the XbaI/EcoRI fragment of -773 TH CAT (-364 to f274, encompassing the TH initiation site at + 1, TH sequences to +27, and 247 NT of CAT sequence) was subcloned into pGEM-1, and linearized at -109 with Bsu36I.…”

Section: Nuclease Protection Assaysmentioning

confidence: 99%

Sequences That Direct Rat Tyrosine Hydroxylase Gene Expression

Fung

Yoon

Chikaraishi

1992

Journal of Neurochemistry

102

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Investigation of neuroendocrine genes has revealed that transcription is regulated via multiple DNA binding sites, including the cyclic AMP response element (CRE). We show here that for the neuronal and chromaffin-specific gene tyrosine hydroxylase (TH), a 70-bp region (-229 to -160) lacking the CRE is sufficient, in either orientation, to confer levels of chloramphenicol acetyltransferase reporter expression equivalent to or greater than that conferred by 4.8 kb of the rat TH enhancer/promoter region. The 70-bp region contains potential binding sites for AP2, AP1, E2A/MyoD, and POU transcription factors, and functions when linked to the TH promoter, but not when joined to a heterologous RSV promoter. This demonstrates that promoter as well as enhancer elements are important for TH expression. In gel-shift assays, the 70-bp fragment forms a cell type-specific complex with nuclear extracts from TH-expressing cells. which is effectively competed by an oligonucleotide containing AP2, AP1, and E2A/MyoD (E box) sites, but not by one containing the POU site. These data suggest that the AP2, AP1, and/or E box sites may be involved in forming the cell-specific complex. Although it lacks an authentic CRE, the 70-bp region also mediated a twofold transcriptional response to forskolin, equivalent to that found with the endogenous gene. A different region (-60 to -29) bearing a consensus CRE mediated a sixfold increase in transcription in response to forskolin, but only minimally activated basal transcription from the TH promoter in the absence of forskolin.

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Brain-specific polyA- transcripts are detected in polyA+ RNA: do complex polyA- brain RNAs really exist?

Cited by 12 publications

References 29 publications

The Cyclic AMP Response Element Directs Tyrosine Hydroxylase Expression in Catecholaminergic Central and Peripheral Nervous System Cell Lines from Transgenic Mice

The Cyclic AMP Response Element Directs Tyrosine Hydroxylase Expression in Catecholaminergic Central and Peripheral Nervous System Cell Lines from Transgenic Mice

Characterization of MurineGirk2Transcript Isoforms: Structure and Differential Expression

Sequences That Direct Rat Tyrosine Hydroxylase Gene Expression

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