Preferential deadenylation of Sarcophaga lectin mRNA during its acute phase expression

Nanbu, Rika; Kubo, Takeo; Ueno, Kensuke; Natori, Shunji

doi:10.1016/0167-4781(91)90154-e

Cited by 1 publication

(1 citation statement)

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“…RNase H Digestions-The method of Nanbu et al (1991) was used. RNA aliquots (15 jLg) were denatured in 80% formamide at 65°C for 10 min and annealed with 15 jLg of oligotd'I') (Pharmacia Biotech Inc.) or with 1 jLg of an oligonucleotide complementary to the SOD•1 mRNA (described below) for 90 min at 50°C in 10 mM Pipes, pH 6.4, 40% formamide, 0.4 MNaCl, and 1 mM EDTA.…”

Section: Methodsmentioning

confidence: 99%

In Male Mouse Germ Cells, Copper-Zinc Superoxide Dismutase Utilizes Alternative Promoters That Produce Multiple Transcripts with Different Translation Potential

Morales

Hecht

1995

Journal of Biological Chemistry

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Copper-zinc superoxide dismutase (SOD-1) is an enzyme that is widely expressed in eukaryotic cells and performs a vital role in protecting cells against free radical damage. In mouse testis, three different sizes of SOD-1 mRNAs of about 0.73, 0.80, and 0.93 kilobases (kb) are detected. The 0.73-kb mRNA is found in early stages of male germ cells and in all somatic tissues. The mRNAs of 0.80 and 0.93 kb are exclusively detected in post-meiotic germ cells. RNase H digestions and Northern blot analyses reveal that the three SOD-1 mRNAs are derived from two transcripts, a ubiquitously expressed transcript and a post-meiotic transcript, which differ by 114-120 nucleotides. RNase protection assays demonstrate that the additional nucleotides present in the post-meiotic mRNA are solely in the 5'-untranslated region. Using a probe derived from the 5'-untranslated region of the 0.93-kb SOD-1 mRNA, we have established that it originates from an alternative upstream promoter contiguous with the somatic SOD-1 promoter. Polysomal gradient analysis of the three mouse testis SOD-1 mRNAs reveals that the 0.93-kb SOD-1 mRNA is primarily non-polysomal, while the 0.80- and 0.73-kb SOD-1 mRNAs are mostly polysome associated. A faster migrating form of the 0.93-kb SOD-1 mRNA is present on polysomes as a result of partial deadenylation. In a cell-free translation system, the 0.73-kb SOD-1 mRNA translates about 2-fold more efficiently than the 0.93-kb SOD-1 mRNA. These data demonstrate that male germ cells transcribe two size classes of SOD-1 mRNAs with different translation potential by utilizing two different promoters, post-meiotic SOD-1 mRNAs undergo adenylation changes, and one of the post-meiotic SOD-1 mRNAs is transcribed during mid-spermiogenesis and translated days later in a partially deadenylated form.

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Section: Methodsmentioning

confidence: 99%