Functional interplay between ribosomal protein paralogues in the eRpL22 family in Drosophila melanogaster
Duplicated ribosomal protein (RP) genes in the Drosophila melanogaster eRpL22 family encode structurally-divergent and differentially-expressed rRNA-binding RPs. eRpL22 is expressed ubiquitously and eRpL22-like expression is tissue-restricted with highest levels in the adult male germline. We explor...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2018-10-01
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| Series: | Fly |
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| Online Access: | http://dx.doi.org/10.1080/19336934.2018.1549419 |
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| author | Catherine M. Mageeney Michael G. Kearse Brett W. Gershman Caroline E. Pritchard Jennifer M. Colquhoun Vassie C. Ware |
| author_facet | Catherine M. Mageeney Michael G. Kearse Brett W. Gershman Caroline E. Pritchard Jennifer M. Colquhoun Vassie C. Ware |
| author_sort | Catherine M. Mageeney |
| collection | DOAJ |
| description | Duplicated ribosomal protein (RP) genes in the Drosophila melanogaster eRpL22 family encode structurally-divergent and differentially-expressed rRNA-binding RPs. eRpL22 is expressed ubiquitously and eRpL22-like expression is tissue-restricted with highest levels in the adult male germline. We explored paralogue functional equivalence using the GAL4-UAS system for paralogue knockdown or overexpression and a conditional eRpL22-like knockout in a heat- shock flippase/FRT line. Ubiquitous eRpL22 knockdown with Actin-GAL4 resulted in embryonic lethality, confirming eRpL22 essentiality. eRpL22-like knockdown (60%) was insufficient to cause lethality; yet, conditional eRpL22-like knockout at one hour following egg deposition caused lethality within each developmental stage. Therefore, each paralogue is essential. Variation in timing of heat-shock-induced eRpL22-like knockout highlighted early embryogenesis as the critical period where eRpL22-like expression (not compensated for by eRpL22) is required for normal development of several organ systems, including testis development and subsequent sperm production. To determine if eRpL22-like can substitute for eRpL22, we used Actin-GAL4 for ubiquitous eRpL22 knockdown and eRpL22-like-FLAG (or FLAG-eRpL22: control) overexpression. Emergence of adults demonstrated that ubiquitous eRpL22-like-FLAG or FLAG-eRpL22 expression eliminates embryonic lethality resulting from eRpL22 depletion. Adults rescued by eRpL22-like-FLAG (but not by FLAG-eRpL22) overexpression had reduced fertility and longevity. We conclude that eRpL22 paralogue roles are not completely interchangeable and include functionally-diverse roles in development and spermatogenesis. Testis-specific paralogue knockdown revealed molecular phenotypes, including increases in eRpL22 protein and mRNA levels following eRpL22-like depletion, implicating a negative crosstalk mechanism regulating eRpL22 expression. Paralogue depletion unmasked mechanisms, yet to be defined that impact paralogue co-expression within germ cells. |
| first_indexed | 2024-03-11T22:58:32Z |
| format | Article |
| id | doaj.art-594021bd57b64e47ba00c53a753a9536 |
| institution | Directory Open Access Journal |
| issn | 1933-6934 1933-6942 |
| language | English |
| last_indexed | 2024-03-11T22:58:32Z |
| publishDate | 2018-10-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Fly |
| spelling | doaj.art-594021bd57b64e47ba00c53a753a95362023-09-21T15:09:08ZengTaylor & Francis GroupFly1933-69341933-69422018-10-01123-414316310.1080/19336934.2018.15494191549419Functional interplay between ribosomal protein paralogues in the eRpL22 family in Drosophila melanogasterCatherine M. Mageeney0Michael G. Kearse1Brett W. Gershman2Caroline E. Pritchard3Jennifer M. Colquhoun4Vassie C. Ware5Lehigh UniversityLehigh UniversityLehigh UniversityLehigh UniversityLehigh UniversityLehigh UniversityDuplicated ribosomal protein (RP) genes in the Drosophila melanogaster eRpL22 family encode structurally-divergent and differentially-expressed rRNA-binding RPs. eRpL22 is expressed ubiquitously and eRpL22-like expression is tissue-restricted with highest levels in the adult male germline. We explored paralogue functional equivalence using the GAL4-UAS system for paralogue knockdown or overexpression and a conditional eRpL22-like knockout in a heat- shock flippase/FRT line. Ubiquitous eRpL22 knockdown with Actin-GAL4 resulted in embryonic lethality, confirming eRpL22 essentiality. eRpL22-like knockdown (60%) was insufficient to cause lethality; yet, conditional eRpL22-like knockout at one hour following egg deposition caused lethality within each developmental stage. Therefore, each paralogue is essential. Variation in timing of heat-shock-induced eRpL22-like knockout highlighted early embryogenesis as the critical period where eRpL22-like expression (not compensated for by eRpL22) is required for normal development of several organ systems, including testis development and subsequent sperm production. To determine if eRpL22-like can substitute for eRpL22, we used Actin-GAL4 for ubiquitous eRpL22 knockdown and eRpL22-like-FLAG (or FLAG-eRpL22: control) overexpression. Emergence of adults demonstrated that ubiquitous eRpL22-like-FLAG or FLAG-eRpL22 expression eliminates embryonic lethality resulting from eRpL22 depletion. Adults rescued by eRpL22-like-FLAG (but not by FLAG-eRpL22) overexpression had reduced fertility and longevity. We conclude that eRpL22 paralogue roles are not completely interchangeable and include functionally-diverse roles in development and spermatogenesis. Testis-specific paralogue knockdown revealed molecular phenotypes, including increases in eRpL22 protein and mRNA levels following eRpL22-like depletion, implicating a negative crosstalk mechanism regulating eRpL22 expression. Paralogue depletion unmasked mechanisms, yet to be defined that impact paralogue co-expression within germ cells.http://dx.doi.org/10.1080/19336934.2018.1549419cell biologyfertilizationgene regulationmolecular geneticsreproductionrnai |
| spellingShingle | Catherine M. Mageeney Michael G. Kearse Brett W. Gershman Caroline E. Pritchard Jennifer M. Colquhoun Vassie C. Ware Functional interplay between ribosomal protein paralogues in the eRpL22 family in Drosophila melanogaster Fly cell biology fertilization gene regulation molecular genetics reproduction rnai |
| title | Functional interplay between ribosomal protein paralogues in the eRpL22 family in Drosophila melanogaster |
| title_full | Functional interplay between ribosomal protein paralogues in the eRpL22 family in Drosophila melanogaster |
| title_fullStr | Functional interplay between ribosomal protein paralogues in the eRpL22 family in Drosophila melanogaster |
| title_full_unstemmed | Functional interplay between ribosomal protein paralogues in the eRpL22 family in Drosophila melanogaster |
| title_short | Functional interplay between ribosomal protein paralogues in the eRpL22 family in Drosophila melanogaster |
| title_sort | functional interplay between ribosomal protein paralogues in the erpl22 family in drosophila melanogaster |
| topic | cell biology fertilization gene regulation molecular genetics reproduction rnai |
| url | http://dx.doi.org/10.1080/19336934.2018.1549419 |
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