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Published on: March 16, 2016
[Protein synthesizing structures and protein synthesis in mouse lymphocytes]
Abstract:
The properties of cytoplasmic mRNP particles from spleen lymphocytes (sedimentation coefficients and buoyant density) and the kinetics of incorporation of impulse amino acid label at definite labelling periods (5, 15, 30, 60 sec) were studied. After 5 or 15 sec labelling periods the labelled amino acid was detected in the polysomes (sedimentation coefficient 130-240S, buoyant density 1.55 g/cm3) and in small-sized particles (sedimentation coefficient 80S and less, buoyant density 1.34, 1.50 and 1.60 g/cm3). After 30 sec of labelling the label is incorporated into the medium-sized particles (sedimentation coefficient 80-120 S, buoyant density 1.47 g/cm3); no increase of the label incorporation into the small-sized particles is observed. After 60 sec the label content in all the three sucrose gradient zones is increased. The observed succession of the amino acid label incorporation first into the small-sized particles and then into the large-sized ones having a different buoyant density and the data from impulse uridyl labelling suggest that in the lymphocytes the translation occurs already in the complexes of informosomes with single ribosomes and continues in the course of their conversion to polysomes. The short-time labelling of the cells with an impulse amino acid label allows to determine the time of the polypeptide chain translation from the time of polysome saturation with the label.
Insights
Messenger ribonucleoprotein (mRNP) particles in spleen lymphocytes were studied using amino acid labeling. Findings suggest translation begins in informosome-ribosome complexes and continues during polysome formation.
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Context:
- Investigates cytoplasmic messenger ribonucleoprotein (mRNP) particles in spleen lymphocytes.
- Examines the kinetics of amino acid incorporation into these particles at early time points (5-60 seconds).
- Characterizes mRNP particle properties including sedimentation coefficients and buoyant density.
Purpose:
- To elucidate the early stages of protein synthesis and polysome assembly in lymphocytes.
- To determine the temporal sequence of amino acid labeling in different mRNP fractions.
- To understand the role of informosomes in initiating translation.
Summary:
- Short-term amino acid labeling revealed sequential incorporation into small (≤80S) and then larger (80-120S and polysomal) mRNP particles.
- Distinct buoyant densities were observed for different particle sizes, indicating compositional differences.
- Data suggest translation initiates on informosome-ribosome complexes and proceeds as these complexes mature into polysomes.
Impact:
- Provides insights into the dynamic process of translation initiation and elongation in lymphocytes.
- Establishes a method using pulse labeling to estimate polypeptide chain synthesis time.
- Contributes to understanding gene expression regulation at the post-transcriptional level.
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